Microspore ganoderma lucidum immune protein and herba houttuyniae fermentation for treating AIDS and application

Abstract

The invention relates to the technical field of traditional Chinese medicine extract fermentation, and provides a microspore ganoderma lucidum immune protein and herba houttuyniae fermentation method for treating AIDS and application of microspore ganoderma lucidum immune protein and herba houttuyniae fermentation method.The microspore ganoderma lucidum immune protein and herba houttuyniae fermentation method comprises the steps that microspore ganoderma lucidum strains and herba houttuyniae fermentation liquor are co-fermented, beta-cyclodextrin-selenocystine of a specially-made additive is combined, and the microspore ganoderma lucidum immune protein is obtained; the problems that in traditional AIDS treatment, latent infection cells are difficult to remove, toxic and side effects of drugs are remarkable, and the reconstruction efficiency of an immune system is low are effectively solved, experiments prove that the fermentation product can activate the immune system, the cell number and the dendritic cell maturity are remarkably improved, and the curative effect is good. Meanwhile, the multi-target antiviral effect is achieved by blocking virus envelope combination, inhibiting reverse transcriptase activity and inducing latent infection cell apoptosis; the anti-oxidation and anti-inflammatory characteristics can reduce the risk of opportunistic infection, reduce the toxic damage of antiviral drugs to liver, kidney and intestinal tracts, and provide a new scheme for comprehensive treatment of AIDS and nutrition support of patients.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine extract fermentation technology, specifically referring to the fermentation and application of microspore Ganoderma lucidum immune protein and Houttuynia cordata for the treatment of AIDS. Background Technology

[0002] Currently, the treatment of AIDS mainly relies on antiretroviral therapy, whose core goal is to inhibit viral replication. Although it can slow disease progression, it cannot eliminate the HIV viral reservoir latent in host cells, and long-term use can easily lead to drug resistance and toxic side effects. Against this backdrop, the multi-target effects and low toxicity of natural medicines have become a research hotspot. Immunomodulatory proteins in Ganoderma lucidum spores have been shown to activate macrophages, promote lymphocyte proliferation, and enhance cytokine secretion, thereby improving the body's immunity. Houttuynia cordata, with its components such as chrysophanol, has antibacterial, anti-inflammatory, and immunomodulatory functions. Both have shown antiviral potential when used alone. However, existing research mostly focuses on the extraction of single components and lacks exploration of compound preparations for the treatment of AIDS.

[0003] The current technical challenges mainly lie in suppressing viral rebound and reducing treatment-related toxicity. Traditional methods are difficult to eliminate latent infected cells, leading to rebound. At the same time, antiviral drugs often cause mitochondrial damage and liver and kidney damage, which exacerbates patient compliance. Microspore Ganoderma lucidum technology is difficult to maintain stability and purity of components in large-scale production. In addition, there is a risk of opportunistic infection during immune reconstitution, requiring a better regulatory mechanism to balance efficacy and safety. Summary of the Invention

[0004] In response to the above situation and to overcome the shortcomings of the existing technology, this invention provides a microspore Ganoderma lucidum immune protein for the treatment of AIDS, fermentation and application of Houttuynia cordata, which effectively solves the problems of traditional AIDS treatment drugs on the market that are unable to clear latent infected cells and thus cause rebound, antiviral drugs that increase patient compliance and are prone to side effects, difficulty in maintaining stability and purity of components, and the risk of opportunistic infections.

[0005] The technical solution adopted in this invention is as follows: This invention proposes a microspore Ganoderma lucidum immune protein for treating AIDS, fermentation of Houttuynia cordata, and its application, including the following steps: A slant culture medium was prepared, sterilized, and then cultured at a constant temperature with *Ganoderma lucidum* spores. After the mycelium covered the slant, it was inoculated into a seed tank to prepare seed liquid. Simultaneously, *Houttuynia cordata* fermentation broth was prepared. Fresh *Houttuynia cordata* was washed, pressed, filtered, and the sugar content was adjusted. After high-temperature sterilization, fermentation was carried out to obtain *Houttuynia cordata* fermentation broth. A fermentation culture medium was prepared, and *Ganoderma lucidum* seed liquid and *Houttuynia cordata* fermentation broth were mixed in proportion for co-fermentation. Carbon source, nitrogen source, and inorganic salts were added, and two-stage fermentation was carried out to obtain fermented product. The fermented product was heated, a complex protease was added, and the pH was controlled for enzymatic hydrolysis. 0.5% β-cyclodextrin-selenocysteine ​​was added and stirred for reaction. After filtration and concentration, *Ganoderma lucidum* immunoprotein and *Houttuynia cordata* fermentation broth were obtained.

[0006] Furthermore, the preparation method of the β-cyclodextrin-selenocysteine ​​includes the following steps: 1,1'-carbonyldiimidazole was added to the β-cyclodextrin solution, and the mixture was stirred in the dark. After the reaction was completed, the precipitate was discarded by centrifugation at low temperature and high speed, and the supernatant was collected. The selenocysteine ​​solution was slowly added dropwise to the activated β-cyclodextrin solution and catalyzed. The reaction solution was purified by dialysis and then freeze-dried to obtain β-cyclodextrin-selenocysteine.

[0007] Furthermore, the fresh houttuynia cordata is washed, pressed, and the juice is filtered and the sugar content is adjusted to 13.5-14.5%, and 0.01% KH3PO4 is added.

[0008] Furthermore, the fermentation process is divided into three stages: the first stage involves inoculating yeast and fermenting at 28°C for 48 days; the second stage involves transferring the fermentation liquid to a Karl von Willebrand tank, adding 0.01% KH3PO4, and fermenting at 25°C until the alcohol content reaches 13.5-14.5%; the third stage involves letting the fermentation liquid stand at 16°C for 25 days, fumigating with sulfur to inhibit bacteria, and filtering to obtain the Houttuynia cordata fermentation liquid.

[0009] Furthermore, the Ganoderma lucidum seed liquid and Houttuynia cordata fermentation liquid are mixed in proportion, with the volume ratio of Ganoderma lucidum seed liquid to Houttuynia cordata fermentation liquid being 1:(2-4).

[0010] Furthermore, the two-stage fermentation is divided into aerobic fermentation and anaerobic static fermentation. The aerobic fermentation conditions are 30°C, a shaking speed of 200 rpm, and 7 days of cultivation to promote mycelial growth and accumulate Ganoderma lucidum polysaccharides. The anaerobic static fermentation conditions are 28°C, sealed and protected from light, for 21 days to induce triterpenoid synthesis and Houttuynia cordata flavonoid conversion.

[0011] Further, 1,1'-carbonyldiimidazole is added to the β-cyclodextrin solution, and the mass ratio of β-cyclodextrin to 1,1'-carbonyldiimidazole is 100:(4-6).

[0012] Furthermore, the catalytic reaction was carried out by adding 0.5 mL of triethylamine as a catalyst and stirring at 40°C in the dark for 8 hours.

[0013] Furthermore, for the purification process, a dialysis bag with a molecular weight cutoff of 3.5 kDa is selected, and the mobile phase used in the purification process is deionized water, which is replaced every 4 hours for a total of 20 hours.

[0014] Furthermore, the microspore Ganoderma lucidum immune protein and Houttuynia cordata fermentation product are used for comprehensive treatment of AIDS and nutritional support food for AIDS patients.

[0015] The beneficial effects achieved by the present invention using the above structure are as follows: This protocol proposes a method for treating AIDS using microspore-derived Ganoderma lucidum immunoglobulin in fermentation and application with Houttuynia cordata. The hydrophobic cavity of β-cyclodextrin encapsulates and protects the Ganoderma lucidum immunoglobulin, preventing damage to its activity from organic acids and enzymes in the fermentation broth. This also prevents denaturation or aggregation and inactivation of the immunoglobulin during preparation and storage. Simultaneously, the hydrophilic groups on the outer edge of the cyclodextrin enhance the water solubility of the complex, overcoming the dissolution barriers caused by the hydrophobicity of biomolecules. This ensures efficient drug dispersion in body fluids, protecting the microspore-derived Ganoderma lucidum immunoglobulin, activating macrophages and T lymphocytes, and enhancing the body's non-specific immune response, particularly effective against HIV-related infections in AIDS patients. Cellular exhaustion and the rebuilding of a damaged immune system can be achieved through the synergistic effect of β-cyclodextrin-selenocysteine ​​and Ganoderma lucidum immune proteins, which together slow down the progression of immune system damage in AIDS.

[0016] The cyclodextrin surface modified with selenocysteine ​​can bind to the DC-SIGN receptor on the surface of dendritic cells to present antigens during HIV immunization. β-cyclodextrin-selenocysteine ​​delivers Ganoderma lucidum immunoproteins to dendritic cells through a receptor-ligand mediated pathway, enhancing drug accumulation in target cells and avoiding waste caused by non-specific distribution. Targeted delivery further activates the antigen-presenting function of dendritic cells, promotes the efficiency of T-cell immune responses, and reduces the rate of viral generation. At the same time, the fermentation process can also enable the active ingredients in Houttuynia cordata to target the HIV latent reservoir, induce the death of latent infected cells, and enhance the body's immune clearance ability against HIV.

[0017] After selenocysteine ​​is metabolized into selenocysteine ​​within cells, it can serve as a cofactor for glutathione peroxidase, enhancing the enzyme's catalytic activity, scavenging excess reactive oxygen species in the HIV infection microenvironment, and blocking the effects of oxidative stress. The cell damage pathway is enhanced; simultaneously, the activity of immune cells is enhanced, selenium uptake and utilization are improved, and the improvement of the oxidative environment further maintains the stability of immune cells, thereby achieving an antiviral mechanism; in addition, microspore Ganoderma lucidum immune proteins promote the activation of macrophages and natural killer cells by activating the Toll-like receptor signaling pathway, enhancing the ability to clear pathogens. Combined with the antibacterial products such as chlorogenic acid and houttuynin in Houttuynia cordata fermentation products, they inhibit the common pathogenic bacteria such as Staphylococcus aureus and Streptococcus pneumoniae, reducing the probability of infection of related diseases. In synergy with β-cyclodextrin-selenocysteine, the antiviral mechanism is enhanced, enabling AIDS patients to avoid complications or inflammation caused by immunodeficiency.

[0018] The cyclodextrin in β-cyclodextrin-selenocysteine ​​slowly dissociates in the blood, preventing premature drug degradation in non-target tissues and significantly prolonging the time for immune proteins to maintain effective blood drug concentrations. Simultaneously, the sustained-release properties of selenocysteine ​​reduce blood drug concentration fluctuations associated with traditional selenium supplements, avoiding concentration-induced toxicity risks. This not only reduces dosing frequency but also ensures the drug maintains an inhibitory concentration at the viral latency site. Furthermore, microspore Ganoderma lucidum immune proteins regulate hepatic cytochrome P450 enzyme activity, accelerating the detoxification and excretion of drug metabolites and reducing the risk of hepatocellular damage. Quercetin produced by Houttuynia cordata fermentation can repair drug-induced intestinal mucosal barrier damage, reducing the incidence of diarrhea and malnutrition. These three factors synergistically maintain treatment tolerability and avoid side effects such as hepatotoxicity and metabolic disorders that are common with long-term antiviral drug treatment, making it suitable for the management of HIV patients requiring long-term medication. Attached Figure Description

[0019] Figure 1 This image shows the results of verifying the immune reconstitution ability of microspore Ganoderma lucidum immune protein and Houttuynia cordata fermentation for treating AIDS, as proposed in this invention.

[0020] Figure 2 The figure shows the experimental results of antiviral activity and latent reservoir clearance of a microspore Ganoderma lucidum immune protein and Houttuynia cordata fermentation proposed in this invention for treating AIDS.

[0021] Figure 3 The figure shows the experimental results of the prevention of complications and the alleviation of drug toxicity of a microspore Ganoderma lucidum immune protein and Houttuynia cordata fermentation proposed in this invention for the treatment of AIDS.

[0022] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used together with the embodiments of the invention to explain the invention and do not constitute a limitation thereof. Detailed Implementation

[0023] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention. Example 1

[0024] A microspore-derived Ganoderma lucidum immune protein for treating AIDS, fermented with Houttuynia cordata and its application First, β-cyclodextrin-selenocysteine ​​was prepared by adding 1,1'-carbonyldiimidazole to a β-cyclodextrin solution at a mass ratio of 100:4. The reaction was carried out under light and stirred. After the reaction was completed, the precipitate was discarded by low-temperature high-speed centrifugation, and the supernatant was collected. The selenocysteine ​​solution was slowly added dropwise to the activated β-cyclodextrin solution, and 0.5 mL of triethylamine was added as a catalyst. The reaction was carried out at 40 °C under light and stirred for 8 hours to allow the SeCys amino group to form an amide bond with the activated β-CD. The reaction solution was purified by dialysis using a 3.5 kDa molecular weight cutoff dialysis bag with deionized water as the mobile phase, which was changed every 4 hours for 20 hours to remove free SeCys and byproducts. After freeze-drying, β-cyclodextrin-selenocysteine ​​was obtained.

[0025] Prepare an agar slant culture medium consisting of 20 g / L potato juice, 20 g / L glucose, 20 g / L agar, and 1 L water, with a natural pH. Sterilize at 115℃ for 20 min. After sterilization, incubate Ganoderma lucidum spores at 25℃ for 20 days. Once the mycelium covers the slant, inoculate it into a seed tank. Simultaneously, prepare a Houttuynia cordata fermentation broth. Wash and press fresh Houttuynia cordata, filter the juice, adjust the sugar concentration to 13.5%, and add 0.01% KH2PO4. After high-temperature sterilization, fermentation begins. Add yeast to the Houttuynia cordata culture medium, mix thoroughly, and incubate at 28℃ for 48 h to obtain the Houttuynia cordata culture broth. Transfer the broth to a Karl flask for further fermentation, adding 0.01% H3PO4 to adjust the sugar concentration to 13.5-14.5%. Continue fermentation until no contaminating microorganisms are detected. Pour the yeast broth into the fermentation tank. The yeast culture was continued in a yeast tank. After fermentation, it was allowed to stand at 16℃ for 25 days. After sulfur fumigation for sterilization, it was filtered to obtain the Houttuynia cordata fermentation broth. A fermentation medium was prepared, and the Ganoderma lucidum seed liquid and Houttuynia cordata fermentation broth were mixed at a volume ratio of 1:2 for co-fermentation. Carbon source, nitrogen source and inorganic salt were added. Aerobic fermentation was carried out at 30℃ and a shaking speed of 200 rpm for 7 days to promote mycelial growth and accumulate Ganoderma lucidum polysaccharides. Then, it was sealed and kept in the dark and anaerobic environment at 28℃ for 21 days to induce triterpene synthesis and Houttuynia cordata flavonoid conversion to obtain the fermentation product. The fermentation product was heated to 50℃, a complex protease was added and the pH was controlled at 7.0 for 4 hours of enzymatic hydrolysis. β-cyclodextrin-selenocysteine ​​was added and stirred. After filtration and concentration, the microspore Ganoderma lucidum immunoprotein and Houttuynia cordata fermentation broth were obtained.

[0026] Prepare 9 parts red ginseng, 11 parts angelica, 9 parts dried ginger, 13 parts arborvitae leaves, 12 parts mugwort leaves, 12 parts platycodon root, 20 parts dried tangerine peel, 40 parts microspore Ganoderma lucidum immune protein and Houttuynia cordata ferment, 7 parts Alisma plantago-aquatica, and 11 parts licorice. Mix the extracts of each herb, add excipients, and make into granules. Example 2

[0027] A microspore-derived Ganoderma lucidum immune protein for treating AIDS, fermented with Houttuynia cordata and its application First, β-cyclodextrin-selenocysteine ​​was prepared by adding 1,1'-carbonyldiimidazole to a β-cyclodextrin solution at a mass ratio of 100:5. The reaction was carried out under light and stirred. After the reaction was completed, the precipitate was discarded by low-temperature high-speed centrifugation, and the supernatant was collected. The selenocysteine ​​solution was slowly added dropwise to the activated β-cyclodextrin solution, and 0.5 mL of triethylamine was added as a catalyst. The reaction was carried out at 40 °C under light and stirred for 8 hours to allow the SeCys amino group to form an amide bond with the activated β-CD. The reaction solution was purified by dialysis using a 3.5 kDa molecular weight cutoff dialysis bag with deionized water as the mobile phase, which was changed every 4 hours for 20 hours to remove free SeCys and byproducts. After freeze-drying, β-cyclodextrin-selenocysteine ​​was obtained.

[0028] Prepare an agar slant culture medium consisting of 20 g / L potato juice, 20 g / L glucose, 20 g / L agar, and 1 L water, with a natural pH. Sterilize at 115℃ for 20 min. After sterilization, incubate Ganoderma lucidum spores at 25℃ for 20 days. Once the mycelium covers the slant, inoculate it into a seed tank. Simultaneously, prepare a Houttuynia cordata fermentation broth. Wash and press fresh Houttuynia cordata, filter the juice, adjust the sugar concentration to 14%, and add 0.01% KH2PO4. After high-temperature sterilization, fermentation begins. Add yeast to the Houttuynia cordata culture medium, mix thoroughly, and incubate at 28℃ for 48 h to obtain the Houttuynia cordata culture broth. Transfer the broth to a Karl flask for further fermentation, adding 0.01% H3PO4 to adjust the sugar concentration to 13.5-14.5%. Continue fermentation until no contaminating microorganisms are detected. Pour the yeast broth into the fermentation tank. The fermentation process continued in the mother tank. After completion, the mixture was allowed to stand at 16℃ for 25 days. After sulfur fumigation for sterilization, the mixture was filtered to obtain the Houttuynia cordata fermentation broth. A fermentation medium was prepared, and the Ganoderma lucidum seed liquid and Houttuynia cordata fermentation broth were mixed at a volume ratio of 1:3 for co-fermentation. Carbon source, nitrogen source, and inorganic salts were added. The mixture was cultured aerobically at 30℃ and a shaking speed of 200 rpm for 7 days to promote mycelial growth and accumulate Ganoderma lucidum polysaccharides. Then, the mixture was cultured in a sealed, light-proof, anaerobic environment at 28℃ for 21 days to induce triterpene synthesis and Houttuynia cordata flavonoid conversion, yielding the fermented product. The fermented product was heated to 50℃, and a complex protease was added while controlling the pH at 7.0 for 4 hours of enzymatic hydrolysis. β-cyclodextrin-selenocysteine ​​was added and the mixture was stirred. After filtration and concentration, the mixture was used to obtain microspore Ganoderma lucidum immunoprotein and Houttuynia cordata fermentation broth.

[0029] Prepare 9 parts red ginseng, 11 parts angelica, 9 parts dried ginger, 13 parts arborvitae leaves, 12 parts mugwort leaves, 12 parts platycodon root, 20 parts dried tangerine peel, 40 parts microspore Ganoderma lucidum immune protein and Houttuynia cordata ferment, 7 parts Alisma plantago-aquatica, and 11 parts licorice. Mix the extracts of each herb, add excipients, and make into granules. Example 3

[0030] A microspore-derived Ganoderma lucidum immune protein for treating AIDS, fermented with Houttuynia cordata and its application First, β-cyclodextrin-selenocysteine ​​was prepared by adding 1,1'-carbonyldiimidazole to a β-cyclodextrin solution at a mass ratio of 100:6. The reaction was carried out under light and stirred. After the reaction was completed, the precipitate was discarded by low-temperature high-speed centrifugation, and the supernatant was collected. The selenocysteine ​​solution was slowly added dropwise to the activated β-cyclodextrin solution, and 0.5 mL of triethylamine was added as a catalyst. The reaction was carried out at 40 °C under light and stirred for 8 hours to allow the SeCys amino group to form an amide bond with the activated β-CD. The reaction solution was purified by dialysis using a 3.5 kDa molecular weight cutoff dialysis bag with deionized water as the mobile phase, which was changed every 4 hours for 20 hours to remove free SeCys and byproducts. After freeze-drying, β-cyclodextrin-selenocysteine ​​was obtained.

[0031] Prepare an agar slant culture medium consisting of 20 g / L potato juice, 20 g / L glucose, 20 g / L agar, and 1 L water, with a natural pH. Sterilize at 115℃ for 20 min. After sterilization, incubate Ganoderma lucidum spores at 25℃ for 20 days. Once the mycelium covers the slant, inoculate it into a seed tank. Simultaneously, prepare Houttuynia cordata fermentation broth. Wash and press fresh Houttuynia cordata, filter the juice, adjust the sugar concentration to 14.5%, and add 0.01% KH2PO4. After high-temperature sterilization, fermentation begins. Add yeast to the Houttuynia cordata culture medium, mix thoroughly, and incubate at 28℃ for 48 h to obtain the Houttuynia cordata culture broth. Transfer the broth to a Karl flask for further fermentation, adding 0.01% H3PO4 to adjust the sugar concentration to 13.5-14.5%. Continue fermentation until no contaminating microorganisms are detected. Pour the yeast broth into the fermentation tank. The yeast culture was continued in a yeast tank. After fermentation, it was allowed to stand at 16℃ for 25 days. After sulfur fumigation for sterilization, it was filtered to obtain the Houttuynia cordata fermentation broth. A fermentation medium was prepared, and the Ganoderma lucidum seed liquid and Houttuynia cordata fermentation broth were mixed at a volume ratio of 1:4 for co-fermentation. Carbon source, nitrogen source and inorganic salt were added. Aerobic fermentation was carried out at 30℃ and a shaking speed of 200 rpm for 7 days to promote mycelial growth and accumulate Ganoderma lucidum polysaccharides. Then, it was sealed and kept in the dark and anaerobic environment at 28℃ for 21 days to induce triterpene synthesis and Houttuynia cordata flavonoid conversion to obtain the fermentation product. The fermentation product was heated to 50℃, a complex protease was added and the pH was controlled at 7.0 for 4 hours of enzymatic hydrolysis. β-cyclodextrin-selenocysteine ​​was added and stirred. After filtration and concentration, the microspore Ganoderma lucidum immunoprotein and Houttuynia cordata fermentation broth were obtained.

[0032] Prepare 9 parts red ginseng, 11 parts angelica, 9 parts dried ginger, 13 parts arborvitae leaves, 12 parts mugwort leaves, 12 parts platycodon root, 20 parts dried tangerine peel, 40 parts microspore Ganoderma lucidum immune protein and Houttuynia cordata ferment, 7 parts Alisma plantago-aquatica, and 11 parts licorice. Mix the extracts of each herb, add excipients, and make into granules.

[0033] Comparative Example 1 Prepare 9 parts red ginseng, 11 parts angelica, 9 parts dried ginger, 13 parts arborvitae leaves, 12 parts mugwort leaves, 12 parts platycodon, 20 parts dried tangerine peel, 8 parts houttuynia cordata, 7 parts alisma plantago-aquatica, and 11 parts licorice. Prepare red ginseng powder by crushing the prescribed amount of red ginseng into powder. Then prepare the extract by decocting the remaining herbs (excluding red ginseng) with water. Decoct twice and combine the two filtrates. Filter the combined filtrate and concentrate it to obtain the extract. Finally, granulate the mixture by adding the prepared red ginseng powder to the extract, adding excipients, mixing, and forming granules.

[0034] Comparative Example 2 Using Ganoderma lucidum, ginseng, fermented cordyceps mycelium powder, and Cordyceps sinensis as basic raw materials, and adding rose petals, broken-cell wall Ganoderma lucidum spore powder, Codonopsis pilosula, and Astragalus membranaceus, the alcohol used for extraction is ethanol. The raw materials of Chinese medicinal materials are weighed according to the prescription, soaked in water, heated and decocted multiple times, filtered and combined, and concentrated to an appropriate amount. After cooling, the concentrate is centrifuged at high speed to remove impurities. The concentrate after impurity removal is added to excipients and the required preparation is made using conventional processes. It is used to prepare health products or medicines for the prevention and treatment of AIDS.

[0035] Comparative Example 3 Prepare the following raw materials: Codonopsis pilosula, pearl grass, honeysuckle, toad skin, sophora flavescens, sophora tonkinensis, oldenlandia diffusa, artemisia capillaris, polygonum cuspidatum, selaginella tamariscina, red peony root, trichosanthes kirilowii, salvia miltiorrhiza, turtle shell, oyster shell, houttuynia cordata, scutellaria barbata, codonopsis pilosula, notoginseng, turmeric, sparganium, morinda officinalis, mulberry mistletoe, rehmannia glutinosa, alisma plantago-aquatica, poria cocos, polyporus umbellatus, centipede, scorpion, and licorice. The proportions of the medicinal materials are equal. The preparation uses a mixture of medicinal material extracts. This preparation can be made into pills, powders, ointments, granules, tablets, capsules, or syrups.

[0036] Experimental Example 1 Immune reconstitution capacity was assessed using a humanized BLT mouse SIV model infected with HIV-1. Ten mice were assigned to each group. The experimental group received the sample reagents from Examples 1-3 via gavage at a daily dose of 200 mg / kg. The control group received the same dose via gavage as in Examples 1-3. The positive control group received tenofovir combined with lamivudine at a daily dose of 50 mg / kg for 8 weeks. During the experiment, the ratio of CD4-positive to CD8-positive T cells in peripheral blood and the expression level of dendritic cell maturity were detected weekly using flow cytometry. Spleen tissue sections were stained to assess the integrity of lymphoid follicle structure. ELISA was used to detect the infiltration degree of CD4-positive cells in spleen tissue. The final efficacy determination required a CD4-positive T cell recovery rate exceeding 30% and a dendritic cell maturity increase reaching twice that of the control group. The experimental results were averaged.

[0037] Table 1 Results of Immune Reconstitution Capacity Validation

[0038] Experimental results are as follows Figure 1 As shown in Table 1, the samples from Examples 1-3 in the experimental group... Cell recovery rate, dendritic cell maturity, serum IL-12 and IFN-γ levels, and spleen In terms of cell infiltration and follicular structural integrity, it was significantly better than the control group (samples 1-3) and the positive control group. Among them, Example 1 showed the best immune improvement effect. The overall data showed a dose-dependent positive trend, demonstrating the significant efficacy of the co-fermentation product in promoting the recovery of immune system function.

[0039] Experimental Example 2 An in vitro cell model was established by infecting MT-4 cells with HIV-1 IIIB strain at a multiplicity of infection (MOI) of 0.01. An in vivo model was established using humanized mice with an integrated proviral DNA reservoir. Examples 1-3 served as the experimental group, with an in vitro concentration of 100 ng / mL and an in vivo dose of 150 mg / kg daily. Comparative examples 1-3 served as the control group, with a daily dose of 200 mg / kg. Nevirapine was used as the positive control at an in vitro concentration of 10 μM / L and an in vivo dose of 30 mg / kg daily. Viral load and reverse transcriptase activity were measured using qRT-PCR. Viral p24 protein expression was analyzed by Western blotting. After stimulation of the latent reservoir, proviral DNA was detected by Alu-PCR, and apoptosis was detected by Annexin V and PI. The key parameter validity threshold was set as a reduction in viral load by more than 2 log units and a latent reservoir cell reduction rate of more than 50% compared to the control group. The experimental results were averaged.

[0040] Table 2. Results of antiviral activity and latent reservoir clearance experiments

[0041] Experimental results are as follows Figure 2 As shown in Table 2, Examples 1-3 demonstrated significant advantages in inhibiting viral replication and clearing latent infected cells. Compared with Comparative Examples 1-3, all Examples 1-3 significantly reduced viral load and effectively cleared latent reservoir cells. Among them, Example 2 showed the best effect. In Comparative Examples 1-3, Comparative Example 1, which contained only unfermented Houttuynia cordata, and Comparative Example 3, which contained multiple medicinal materials, showed some antiviral activity, but the effect was lower than that of Examples 1-3. Comparative Example 2 showed the weakest antiviral ability. The experimental results indicate that the microspore Ganoderma lucidum immunoglobulin and the Houttuynia cordata fermentation process play an important role in enhancing anti-HIV activity.

[0042] Experimental Example 3 First, an HIV-infected mouse model was established, with HIV infection complicated by Staphylococcus aureus. Then, mice were administered a combination of microspore Ganoderma lucidum immunoglobulin and Houttuynia cordata fermentation product with antiviral drugs. The experimental group was treated with the sample reagents from Examples 1-3 via gavage at a daily dose of 200 mg / kg. The control group received the same dose via gavage as in Examples 1-3. The positive control group was treated with tenofovir combined with lamivudine at a daily dose of 50 mg / kg for 8 weeks. After treatment, infection control indicators, oxidative stress indicators, and liver and kidney function indicators were measured. Intestinal barrier function was detected by immunofluorescence, and fecal calprotectin levels were measured. Finally, the effectiveness was evaluated, requiring a reduction in infection mortality of at least 40% and a decrease in hepatobiliary injury markers of at least 30%. The results were compared with those of the antiviral drug-only group, and the average value was taken.

[0043] Table 3. Results of experiments on complication prevention and drug toxicity mitigation

[0044] Experimental results are as follows Figure 3 As shown in Table 3, the combined treatment in Examples 1-3 significantly reduced the mortality rate of the co-infection model, effectively inhibited the proliferation of pathogenic microorganisms in tissues, and significantly improved the phagocytic function of neutrophils. At the same time, it showed significant advantages in reducing oxidative stress damage, maintaining the stability of liver function indicators, and protecting the integrity of the intestinal barrier. Comparative Examples 1-3 showed weaker effects in infection control and organ protection. The experimental results indicate that the synergistic effect of microspore Ganoderma lucidum immunoglobulin and Houttuynia cordata fermentation has a positive effect on the prevention of AIDS complications and the relief of drug toxicity.

[0045] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0046] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

[0047] The present invention and its embodiments have been described above. This description is not restrictive, and the accompanying drawings are only one embodiment of the present invention; the actual structure is not limited thereto. In conclusion, if those skilled in the art are inspired by this description and design similar structures and embodiments without departing from the spirit of the invention, such designs should fall within the protection scope of the present invention.

Claims

1. A microspore-derived Ganoderma lucidum immune protein for treating AIDS, fermented with Houttuynia cordata, characterized in that: The preparation method includes the following steps: A slant culture medium was prepared, sterilized, and then cultured at a constant temperature with *Ganoderma lucidum* spores. After the mycelium covered the slant, it was inoculated into a seed tank to prepare seed liquid. Simultaneously, *Houttuynia cordata* fermentation broth was prepared. Fresh *Houttuynia cordata* was washed, pressed, filtered, and the sugar content was adjusted. After high-temperature sterilization, fermentation was carried out to obtain *Houttuynia cordata* fermentation broth. A fermentation culture medium was prepared, and *Ganoderma lucidum* seed liquid and *Houttuynia cordata* fermentation broth were mixed in proportion for co-fermentation. Carbon source, nitrogen source, and inorganic salts were added, and two-stage fermentation was carried out to obtain fermented product. The fermented product was heated, a complex protease was added, and the pH was controlled for enzymatic hydrolysis. 0.5% β-cyclodextrin-selenocysteine ​​was added and stirred for reaction. After filtration and concentration, *Ganoderma lucidum* immunoprotein and *Houttuynia cordata* fermentation broth were obtained.

2. The microspore Ganoderma lucidum immune protein for treating AIDS and Houttuynia cordata fermentation according to claim 1, characterized in that: The preparation method of the β-cyclodextrin-selenocysteine ​​includes the following steps: 1,1'-carbonyldiimidazole was added to the β-cyclodextrin solution, and the mixture was stirred in the dark. After the reaction was completed, the precipitate was discarded by centrifugation at low temperature and high speed, and the supernatant was collected. The selenocysteine ​​solution was slowly added dropwise to the activated β-cyclodextrin solution and catalyzed. The reaction solution was purified by dialysis and then freeze-dried to obtain β-cyclodextrin-selenocysteine.

3. The microspore Ganoderma lucidum immune protein for treating AIDS and Houttuynia cordata fermentation according to claim 2, characterized in that: The fresh houttuynia cordata is washed, pressed, and filtered. The sugar content is adjusted to 13.5-14.5%, and 0.01% KH3PO4 is added.

4. The microspore Ganoderma lucidum immune protein for treating AIDS and Houttuynia cordata fermentation according to claim 3, characterized in that: The fermentation process is divided into three stages: the first stage involves inoculating yeast and fermenting at 28°C for 48 days; the second stage involves transferring the fermentation liquid to a Karl von Willebrand tank, adding 0.01% KH3PO4, and fermenting at 25°C until the alcohol content reaches 13.5-14.5%; the third stage involves letting the fermentation liquid stand at 16°C for 25 days, fumigating with sulfur to inhibit bacteria, and filtering to obtain the Houttuynia cordata fermentation liquid.

5. The microspore Ganoderma lucidum immune protein for treating AIDS and Houttuynia cordata fermentation according to claim 4, characterized in that: The Ganoderma lucidum seed liquid and Houttuynia cordata fermentation liquid are mixed in proportion, with a volume ratio of 1:(2-4).

6. The microspore Ganoderma lucidum immune protein for treating AIDS and Houttuynia cordata fermentation according to claim 5, characterized in that: The two-stage fermentation is divided into aerobic fermentation and anaerobic static fermentation. The aerobic fermentation conditions are 30°C, a shaking speed of 200 rpm, and 7 days of cultivation. The anaerobic static fermentation conditions are 28°C, sealed and protected from light, and 21 days of cultivation.

7. The microspore Ganoderma lucidum immune protein for treating AIDS and Houttuynia cordata fermentation according to claim 6, characterized in that: 1,1'-carbonyldiimidazole was added to the β-cyclodextrin solution, and the mass ratio of β-cyclodextrin to 1,1'-carbonyldiimidazole was 100:(4-6).

8. The microspore Ganoderma lucidum immune protein for treating AIDS and Houttuynia cordata fermentation according to claim 7, characterized in that: The catalytic reaction was carried out by adding 0.5 mL of triethylamine as a catalyst and stirring at 40 °C in the dark for 8 h.

9. The microspore Ganoderma lucidum immune protein for treating AIDS and Houttuynia cordata fermentation according to claim 8, characterized in that: The dialysis bag used for purification has a molecular weight cutoff of 3.5 kDa. The mobile phase used in the purification process is deionized water, which is replaced every 4 hours for a total of 20 hours.

10. The application of any one of the microspore Ganoderma lucidum immune proteins for treating AIDS according to claims 1-9, and the fermentation of Houttuynia cordata, characterized in that: The microspore Ganoderma lucidum immune protein and Houttuynia cordata fermentation product are used for comprehensive treatment of AIDS and nutritional support food for AIDS patients.

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