Application of Huashibaidu granules for treating novel coronavirus infection in the preparation of drugs for treating porcine blue ear disease

Abstract

The present invention relates to the application of Huashibaidu granules for treating novel coronavirus infection in the preparation of drugs for treating blue ear disease in pigs; wherein, the effective ingredients contained in each 1000g of the Huashibaidu granules are made of the following raw materials in percentage by weight: 14-20% of patchouli, 14-20% of gypsum, 7-10% of magnolia bark, 7-10% of atractylodes, 7-10% of tuckahoe, 4-6% of pinellia tuber, 3-4% of ephedra, 4-6% of stir-fried bitter almond, 5-7% of tsaoko, 2-3% of raw rhubarb, 5-7% of red peony root, 5-7% of scutellaria baicalensis, 5-7% of astragalus, and 1-2% of liquorice. The Huashibaidu granules described in the present application can significantly improve blue ear disease caused by porcine reproductive and respiratory syndrome virus, reduce lung lesions and damage at the macroscopic and microscopic levels, significantly reduce lung viral load, reduce lung enlargement and thymus atrophy, and have a significant effect in treating blue ear disease in pigs.

Description

Technical Field

[0001] The present invention relates to a medical preparation, in particular to a drug preparation containing an undetermined structure from traditional Chinese medicine, and in particular to a Chinese medicine composition for treating blue ear disease in pigs. Background Art

[0002] Porcine reproductive and respiratory syndrome is an acute, contagious viral disease caused by porcine reproductive and respiratory syndrome virus, characterized by reproductive disorders and respiratory symptoms. Infected pigs will have bilateral cyanosis, which is blue-purple, so it is also called blue ear disease. Porcine reproductive and respiratory syndrome virus is an enveloped single-stranded positive-sense RNA virus belonging to the order Nidovirales, the family Arteriviridae, and the genus Arterivirus. The virus particles are approximately spherical or elliptical particles with a diameter of 50-60 nanometers and a relatively smooth appearance.

[0003] The blue ear disease virus is classified as a Class II infectious disease in my country. Pregnant sows and piglets are most susceptible to the disease. Pregnant sows may suffer from miscarriage, stillbirth, weak fetuses and mummified fetuses; boars often have sperm motility, decreased desire, and inflammation of the reproductive system; piglets have poor resistance, and the mortality rate of infected piglets is over 80%; the mortality rate of newly weaned infected pigs is over 50%, and the surviving pigs often become dead pigs. The disease poses a serious threat to the reproduction and survival of pigs, and has caused serious economic losses to the development of the pig farming industry.

[0004] The main clinical symptoms of blue ear disease are cyanosis of both ears, asthma, cough, high fever, lack of energy, loss of appetite, or vomiting, dry stools, conjunctival inflammation, and swollen eyelids.

[0005] At present, inactivated vaccines and attenuated live vaccines are mainly used for prevention. Vaccines can be effective to a certain extent. However, immunosuppression caused by virus mutation and antibody-dependent enhancement of the virus often makes the vaccine ineffective, resulting in immune failure. There is no specific drug for the disease. Generally, antipyretics, analgesics, antibiotics, sulfonamides and antiviral drugs are used as Western medicine treatments. However, the effect is not obvious. If the drugs are used incorrectly or abused, they can easily cause toxic side effects and drug residues in the body of pigs. In addition, Announcement No. 560 of the Ministry of Agriculture and Rural Affairs of the People's Republic of China announced the first batch of "Catalogue of Abolition of Local Standards for Veterinary Drugs", which proposed that some broad-spectrum antiviral drugs (ribavirin, adamantane, etc.) should not be used as veterinary drugs, resulting in few drug choices.

[0006] Qingwen Baidu powder is a famous prescription in traditional Chinese veterinary medicine, which is included in the "Veterinary Pharmacopoeia of the People's Republic of China, Part II". Qingwen Baidu powder is made of 14 herbs: gypsum, rehmannia root, buffalo horn, coptis root, gardenia, peony bark, scutellaria, red peony root, scrophularia, anemarrhena, forsythia, platycodon, liquorice and lophatherum gracile. It has the effects of purging fire, detoxifying and cooling blood, and is mainly used to treat heat toxic spots, high fever and coma. This prescription is a medicine for purging fire from the twelve meridians. Its medicinal composition is mainly composed of cold and cool drugs, which are easy to damage yin, and lack drugs that can remove dampness and replenish the body. After analyzing the pathogenesis and symptoms of blue ear disease, some scholars believe that Qingwen Baidu powder may have the ability to treat blue ear disease in pigs (Zheng Jie et al., Occurrence and prevalence of porcine reproductive and respiratory syndrome caused by highly pathogenic variants in my country, Chinese Journal of Veterinary Medicine, 2007).

[0007] The patent application with publication number CN 116077578 A also discloses a Chinese medicine composition for treating blue ear disease in pigs, which is made of 13-28 parts of scutellaria baicalensis, 12-28 parts of rhubarb, 1-13 parts of gypsum, 6-22 parts of houttuynia, 10-28 parts of mint, 2-16 parts of atractylodes, 5-22 parts of dried orange peel, 5-20 parts of tuckahoe, 4-22 parts of magnolia bark, 3-19 parts of kudzu root, 1-14 parts of glossy privet, 1-13 parts of ephedra, 5-20 parts of apricot kernel, 5-18 parts of gynostemma pentaphyllum, 4-20 parts of astragalus, 2-15 parts of psoralea corylifolia and 1-11 parts of liquorice. The patent formula has as many as 18 flavors, and the formula is complex; the medication cycle is as long as 14 days; at the same time, the antiviral effect in vivo has not been studied.

[0008] In particular, according to the research of the present inventors, the effect of the Chinese medicine composition disclosed in the above patent application in treating blue ear disease in pigs is not ideal. Therefore, it is necessary to explore a treatment plan with stable and reliable efficacy.

[0009] In May 2020, the 5th issue of Volume 22 of Modern Chinese Medicine published an article written by scholar Yang Xiuwei entitled "Research on the Basic Material of the Medicinal Flavor of Huashi Baidu Granules for Pneumonia Caused by New Coronavirus Infection (COVID-19)". The article disclosed that Huashi Baidu Granules had significant effects on the negative conversion of nucleic acid of the new coronavirus (2019-nCoV or SARS-CoV-2) and the improvement of patient symptoms. The above article also disclosed the following content: "Huashi Baidu Prescription" is included in the Diagnosis and Treatment Plan for Pneumonia Caused by New Coronavirus Infection (Trial Seventh Edition), and is listed as a recommended prescription for severe patients with epidemic toxin-blocked lung syndrome. The basic formula of the recommended prescription is 6g of ephedra, 9g of fried bitter almond, 15g of raw gypsum (decoction first), 3g of licorice, 10g of patchouli (added later), 10g of magnolia bark, 15g of atractylodes, 10g of tsaoko, 9g of pinellia tuber, 15g of tuckahoe, 5g of raw rhubarb (added later), 10g of astragalus, 10g of scutellaria, and 10g of red peony root. The dosage is 1 to 2 doses per day, decocted in water, 100mL to 200mL each time, 2 to 4 times a day, orally or by nasogastric feeding. Furthermore, the Diagnosis and Treatment Program for Pneumonia Caused by New Coronavirus Infection (Trial Version 10) also lists the "Huashi Baiduo Fang" as a recommended prescription for epidemic-toxin-blocked lung syndrome in severe new coronavirus infection. The clinical manifestations of epidemic-toxin-blocked lung syndrome are: fever, shortness of breath, chest tightness, cough, yellow and sticky sputum, or blood in sputum, wheezing, dry and bitter mouth, constipation, and short and red urine. The tongue is red, the tongue coating is yellow and greasy, and the pulse is slippery and rapid.

[0010] The patent application with publication number CN 112043804 A discloses a kind of Huashibaidu granules and its preparation method and antiviral drugs. The formula of the drug is the same as the formula of Huashibaidu granules disclosed in the above-mentioned article by Yang Xiuwei, except that the dosage of each medicine is slightly different. At the same time, the description of the patent application also discloses the following content: Huashibaidu granules of the present invention are particularly suitable for treating patients with 2019 novel coronavirus. Preliminary clinical observations show that the present invention can improve the clinical symptoms of pneumonia caused by severe novel coronavirus infection. For severe patients, it can significantly relieve the main symptoms such as cough, fatigue, dry mouth or vomiting. After combined treatment of traditional Chinese and Western medicine, the healing time is shortened, the patient's respiratory function is significantly improved, and the time of oxygen withdrawal is shortened. For ordinary patients, the fever symptoms are significantly relieved, and the symptoms of loss of appetite and chest tightness can also be improved. The clinical symptoms of cough, fatigue, dry mouth or vomiting of pneumonia caused by severe and ordinary novel coronavirus infection are significantly improved, which supplements the treatment of pneumonia caused by severe and ordinary novel coronavirus infection.

[0011] However, since the porcine reproductive and respiratory syndrome virus and the novel coronavirus belong to different families and have completely different nucleic acid sequences, it is impossible to infer that the above-mentioned "Huashi Baidu Fang" is also suitable for the treatment of porcine reproductive and respiratory syndrome. At the same time, there is no relevant report in the existing patent application documents and related journals. Summary of the invention

[0012] The technical problem to be solved by the present invention is to provide a new use of "Huashibaidu Granules" in pharmaceutical manufacturing.

[0013] The new uses of the above-mentioned "Huashi Baidu Granules" in pharmaceutical manufacturing are:

[0014] The use of Huashituidu granules for treating novel coronavirus infection in the preparation of drugs for treating porcine blue ear disease; wherein the effective ingredients contained in every 1000g of the Huashituidu granules are made of the following raw materials in percentage by weight: 14-20% patchouli, 14-20% gypsum, 7-10% magnolia bark, 7-10% atractylodes, 7-10% poria, 4-6% pinellia, 3-4% ephedra, 4-6% stir-fried bitter almonds, 5-7% tsaoko, 2-3% raw rhubarb, 5-7% red peony root, 5-7% scutellaria seed, 5-7% astragalus, and 1-2% licorice.

[0015] In the above application, the preferred embodiment of the active ingredient is made of the following raw materials in percentage by weight: 17.0% patchouli, 17.0% gypsum, 8.5% magnolia bark, 8.5% atractylodes, 8.5% tuckahoe, 5.1% pinellia, 3.4% ephedra, 5.1% stir-fried bitter almonds, 5.6% tsaoko, 2.8% raw rhubarb, 5.6% red peony root, 5.6% scutellaria, 5.6% astragalus, and 1.7% licorice.

[0016] In the above application, the active ingredient is prepared by the following method:

[0017] Take gypsum and add 10 times the amount of water to boil for 20 minutes, then add ephedra, fried bitter almonds, licorice, Magnolia officinalis, Atractylodes macrocephala, Amomum villosum, Radix Paeoniae Alba, Poria, Astragalus, Lepidium japonicus and red peony root and boil for 30 minutes, then add patchouli and rhubarb and boil for 10 minutes, filter to obtain the first decoction; add 8 times the amount of water to the residue and boil for 40 minutes, filter to obtain the second decoction; combine the two decoctions, and concentrate under reduced pressure at 65°C to obtain an extract with a relative density of 1.06-1.20 at 25°C, so as to obtain the active ingredient.

[0018] In the above scheme, the Huashibaidu granules are prepared by the following method:

[0019] Take the extract prepared by the above method, add appropriate amounts of microcrystalline cellulose, soluble starch, sucralose, anhydrous glucose and stevioside, and then use 95% ethanol as a wetting agent to perform dry granulation to obtain the Huashi Baidu granules.

[0020] The formula of the Huashi Baidu Granule of the present invention is composed of 14 ingredients, including patchouli, raw gypsum, Magnolia officinalis, Atractylodes macrocephala, French Pinellia tuber, Poria cocos, Ephedra sinica, stir-fried bitter almond, Lemongrass seed, raw rhubarb, red peony root, Amomum villosum, Astragalus membranaceus and liquorice. In the formula, patchouli and gypsum are the main ingredients. Patchouli is pungent and warm, and can disperse and move qi. It has a fragrant smell and can transform dampness stagnation, dispel filth and harmonize the middle, which is just in line with the pathogenesis of dampness. Gypsum is pungent, sweet and very cold, and can clear away lung heat to produce body fluid. Very cold medicine is used to reduce fever. Magnolia officinalis, Atractylodes macrocephala, Poria cocos and Pinellia tuber are all assistant medicines. Magnolia officinalis, Atractylodes macrocephala and Pinellia tuber are pungent, bitter and warm, and can disperse and move qi. They are bitter and dry in nature and can dry dampness. The three medicines are complementary to each other in that they can dry dampness and strengthen the spleen, promote qi and eliminate fullness. In addition, Poria cocos is sweet, light and flat, and can promote diuresis and eliminate dampness, strengthen the spleen and promote digestion. The four medicines are combined to help the main medicine to eliminate dampness and harmonize the middle. Ephedra, bitter almond, tsaoko, rhubarb, red peony root, scutellaria baicalensis and astragalus are adjuvants. Ephedra is pungent, slightly bitter and warm, and can relieve exterior symptoms, promote diuresis, and relieve asthma. Bitter almond is bitter and slightly warm, and can descend qi, relieve cough and relieve asthma. Scutellaria baicalensis is pungent, bitter and very cold, and can clear the lungs and relieve asthma, promote diuresis and reduce swelling. The three are matched to promote descending and promote mutual influence, and can regulate qi to relieve asthma and promote diuresis. Rhubarb and red peony root are bitter and cold, and can clear heat and remove blood stasis, and conduct heat downward. Tsaoko is pungent and warm, has a fragrant aroma and is good at removing phlegm and drying dampness. Astragalus is sweet and slightly warm, and can replenish qi and raise yang, remove stagnation and relieve arthritis, and treat the vital energy damaged by epidemic diseases, and strengthen the body to encourage the evil to expel. Qi moves and blood circulates, and blood circulates to nourish qi. The seven medicines work together to regulate qi, remove blood stasis and relieve arthritis, promote dampness and eliminate phlegm, so as to help the monarch and minister to transform dampness, and relieve heat and relieve arthritis. Licorice is sweet and flat, can replenish qi and harmonize the middle, clear away heat and detoxify, and harmonize all the medicines as the guiding medicine. The whole formula has the effects of clearing away heat and dampness, drying dampness and strengthening the spleen, relieving Qi, relieving heat and relieving numbness, and strengthening the body and eliminating evil.

[0021] Academician Tong Xiaolin and many famous experts analyzed the etiology and pathogenesis of pneumonia caused by the new coronavirus infection and classified it as "cold and dampness epidemic" (Tong Xiaolin, Exploring the TCM prevention and treatment strategies of pneumonia caused by the new coronavirus infection from the perspective of "cold and dampness epidemic", Journal of Traditional Chinese Medicine, 2020); while Chinese veterinary medicine classified blue ear disease as a febrile disease, and the treatment methods of the two are different. The syndrome differentiation of blue ear disease believes that the disease is caused by the invasion of external evil and the body's inability to resist the disease. It is mainly caused by the continuous accumulation of external moisture in the body, which leads to the stagnation of lung qi, and then the imbalance of yin and yang, resulting in the disease. In traditional Chinese medicine treatment, the main response strategies are "clearing heat and detoxification", "removing dampness" and "moistening the lungs and relieving cough". Therefore, the present invention uses patchouli and gypsum as the main drugs, and makes heavy use of patchouli to aromatically transform dampness, which is in line with the pathogenesis of dampness-heavy disease; making heavy use of gypsum can reduce fever and treat febrile diseases, and secondly, because the properties of dampness-transforming drugs are mostly warm, the properties of very cold drugs are restrained; Magnolia bark, Atractylodes lancea, Poria cocos, and Pinellia are used as assistants to transform dampness, and the dosage of Magnolia bark is increased to further increase the effect of transforming dampness; Ephedra, bitter almond, Amomum villosum, Rhubarb, Red Peony Root, Lemongrass Seed, and Astragalus are used as adjuvants to regulate Qi, relieve heat and relieve numbness, and eliminate dampness and expectorant, so as to help the main and assistant drugs to transform dampness and relieve heat; and licorice is used to harmonize the various drugs, so that the prescription can well comply with the treatment principles of blue ear disease.

[0022] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0023] The Hua Shi Baidu Granules described in the present invention can significantly improve blue ear disease caused by porcine reproductive and respiratory syndrome virus, reduce lung lesions and damage at the macroscopic and microscopic levels, significantly reduce lung viral load, reduce lung enlargement and thymus atrophy, and have a significant effect in treating blue ear disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a curve chart showing the changes in body temperature of different experimental groups during the experimental period.

[0025] Figure 2 Figure 2 shows the organ index (left: lung; right: thymus). The codes #, ## and ### respectively represent the difference between the control group and the model group. # P<0.05, ## P<0.01, ### P<0.001; ns means no statistical significance compared with the model group. The code * means the difference between the experimental drug group and the experimental drug group. * P<0.05, ** P<0.01.

[0026] Figure 3 This is a micrograph of HE sections of lung tissue.

[0027] Figure 4 The histogram of the mRNA expression of porcine reproductive and respiratory syndrome virus NSP9 in different experimental groups, the code nd in the figure represents not detected; # represents compared with the model group, # P<0.05; ns means no statistical significance compared with the model group; code * means compared with the experimental drug group * P<0.05, ** P<0.01. DETAILED DESCRIPTION

[0028] The present invention is described in detail by the following specific examples.

[0029] Example 1 (Particle Preparation Example)

[0030] Example 1

[0031] 1. Prescription:

[0032] Patchouli 300g, gypsum 300g, Magnolia officinalis 150g, Atractylodes 150g, Poria 150g, Pinellia 90g, Ephedra 60g, stir-fried bitter almond 90g, Amomum villosum 100g, raw rhubarb 50g, red peony root 100g, Leonurus japonicus 100g, Astragalus 100g, and licorice 30g.

[0033] 2. Preparation method:

[0034] (1) Add 10 times the amount of water to raw gypsum, boil for 20 minutes, then add ephedra, stir-fried bitter almond, liquorice, magnolia bark, atractylodes, tsaoko, pinellia tuber, tuckahoe, astragalus, scutellaria baicalensis and red peony root, boil for 30 minutes, then add patchouli and rhubarb, boil for 10 minutes, filter, and obtain the first decoction; add 8 times the amount of water to the residue, boil for 40 minutes, filter, and obtain the second decoction; combine the two decoctions, and concentrate under reduced pressure at 65°C to obtain an extract with a density of 1.06 to 1.20 at 25°C;

[0035] (2) Take the extract prepared in step (1), add 467 g of microcrystalline cellulose, 93 g of soluble starch, 60 g of sucralose, 30 g of anhydrous glucose and 30 g of steviol glycoside, and then use 95% ethanol as a wetting agent to perform dry granulation to obtain 1000 g of Huashi Baidu Granules.

[0036] Example 2

[0037] 1. Prescription:

[0038] Patchouli 340g, gypsum 323g, Magnolia officinalis 136g, Atractylodes 136g, Poria 120g, Pinellia 85g, Ephedra 68g, stir-fried bitter almond 103g, Amomum villosum 102g, raw rhubarb 51g, red peony root 85g, Lemongrass seed 85g, Astragalus 102g, and licorice 34g.

[0039] 2. Preparation method: Same as Example 1.

[0040] Example 3

[0041] 1. Prescription:

[0042] Patchouli 240g, gypsum 326g, Magnolia officinalis 173g, Atractylodes 155g, Poria 137g, Pinellia 86g, Ephedra 51g, stir-fried bitter almond 86g, Amomum villosum 103g, raw rhubarb 52g, red peony root 120g, Leonurus japonicus 103g, Astragalus 103g, and licorice 35g.

[0043] 2. Preparation method: Same as Example 1.

[0044] Example 2 (Efficacy Experiment)

[0045] 1. Experimental Drugs

[0046] Test drug: the granules of Example 1 in the above Example 1 (granule preparation example).

[0047] Control drug 1: (i.e., granules prepared from the original Huashi Baidu Fang) is composed of 100g of Patchouli, 150g of Gypsum, 100g of Magnolia officinalis, 150g of Atractylodes macrocephala, 150g of Poria cocos, 90g of Pinellia ternata, 60g of Ephedra, 90g of Stir-fried Bitter Apricot Kernel, 100g of Amomum villosum, 50g of Raw Rhubarb, 100g of Red Peony Root, 100g of Lemongrass Seed, 100g of Astragalus, and 30g of Licorice. Its preparation method is the same as that of test drug 1.

[0048] Control drug 2: Qingwen Baidu San, a commercially available veterinary preparation purchased from Hefei Zhonglong Shenli Animal Pharmaceutical Co., Ltd. (Wei Qianfang), batch number 20230719.

[0049] Control drug 3: Granules were prepared according to the prescription and preparation method of Example 3 of the patent application with publication number CN 116077578 A, and the specific prescription amount was: Scutellaria 18g, Anemarrhena 18g, Rhubarb 6g, Gypsum 18g, Houttuynia cordata 12g, Mentha 18g, Atractylodes 8g, Tangerine peel 12g, Poria 10g, Magnolia officinalis 12g, Pueraria lobata 10g, Ligustrum lucidum 6g, Ephedra 8g, Apricot kernel 10g, Gynostemma pentaphyllum 10g, Astragalus 12g, Psoralea corylifolia 8g, and Licorice 4g.

[0050] 2. Experimental Animals

[0051] 90 Landrace dual-purpose pigs, 4 weeks old, with an average body weight of 6 kg, half male and half female, were randomly divided into 6 groups: blank group, experimental drug group, control drug group 1, control drug group 2, control drug group 3 and model group, with 15 pigs in each group.

[0052] 3. Establishment of porcine reproductive and respiratory syndrome model

[0053] Determination of tissue culture infectious dose (TCID 50 ), diluted to 10 with DMEM medium 5 / mL, for the pigs in the model group and the treatment group, the model was established by 2mL posterior cervical muscle injection + 2mL nasal drops.

[0054] After 24 hours, secretions accumulated in the corners of the pigs' eyes and their eyelids became red and swollen. 3-4 mL of blood was collected from the anterior vena cava of each pig and placed in a sodium heparin blood collection tube. Plasma was obtained by centrifugation and tested by qRT-PCR. The infected pigs were found to be carrying the blue ear disease virus and the model was successfully established.

[0055] 4. Test methods

[0056] Each group was given the same amount of feed, the test drug group was given at a dose of 4.73 g crude drug / kg body weight; the control drug group 1 was given at a dose of 3.66 g crude drug / kg body weight; the control drug group 2 was given at 4 g crude drug / kg body weight; the control drug group 3 was given according to the dose of Example 2 of CN 116077578 A. After mixing the drug with the feed, each drug group was free to eat, while the blank group and the model group were not given any drug and were free to drink water.

[0057] After 7 days of administration, the administration was stopped and the animals were observed for 7 days. The temperature was measured every day during the experiment. After the observation on the 14th day, the pigs were killed and dissected to examine the tissue lesions macroscopically.

[0058] Lung and thymus tissues were removed, and the organ index was calculated using the formula:

[0059] Organ index (%) = organ weight (g) / pig weight (g) * 100%

[0060] Lung tissue was collected for histopathological examination and viral load determination.

[0061] 5. Histopathological Examination

[0062] The tissues at the left hilum were fixed with 4% paraformaldehyde, dehydrated with gradient ethanol, infiltrated, embedded in paraffin, and cut into 5 μm slices. Appropriate slices were selected for observation under a microscope and stained with H&E (hematoxylin & eosin). Histopathological changes were observed under an optical microscope and photographed.

[0063] 6. Lung viral load detection

[0064] The tissues from the left hilum were obtained and homogenized. Total RNA was extracted by TRIZOL method and then determined by fluorescence quantitative PCR.

[0065] The primer designs are shown in Table 1.

[0066] Table 1 Primer sequence list

[0067]

[0068] The reverse transcription reaction system is shown in Table 2.

[0069] Table 2 Reverse transcription reaction system preparation table

[0070]

[0071] The fluorescence quantitative PCR reaction system is shown in Table 3.

[0072]

[0073] Two-step fluorescence quantitative PCR reaction: pre-denaturation at 95°C for 30 s; denaturation at 95°C for 15 s, annealing and extension at 60°C for 30 s, 40 cycles, after the extension ends, collect fluorescence signals; finally, make a melting curve during the process of 65°C - 95°C.

[0074] 7. Statistical analysis

[0075] One-way ANOVA was performed using Graphpad prism 8.0 software, and the data were expressed as mean ± SD.

[0076] 8. Results of pharmacodynamic experiments

[0077] 8.1 Clinical symptoms

[0078] The pigs challenged with the virus showed symptoms such as fever, cough, sneezing, wheezing, and red and swollen eyelids. After drug treatment, the symptoms improved. After dissection, there was no lung injury in the blank group; obvious congestion, swelling, marble-like lesions of red and white alternation, and rubbery lungs were visible on the surface of the lungs in the model group. After drug administration, the lung injuries in the experimental drug group, control drug 1 group, control drug 2 group, and control drug 3 group were improved. Among them, the effect of the experimental drug in improving the lesions was better than that of control drug 1, control drug 2, and control drug 3.

[0079] 8.2 Body temperature changes

[0080] The body temperature change graph during the experimental period is as Figure 1 shown, and the body temperature at the end of the experimental period is shown in Table 4. Defining 40°C and above as fever, at the end of the experimental period, the body temperatures of the pigs in the blank group and the drug administration group were within the normal range (below 40°C), while the model group was still in a fever state (above 40°C). After drug treatment, there were statistical differences in the decrease in body temperature between the experimental drug group (P < 0.0001), control drug 1 group (P < 0.0001), control drug 2 group (P < 0.0001), control drug 3 group (P < 0.0001) and the model group. Compared with the experimental drug group, there were statistical differences in the control drug 1 group (P < 0.05), control drug 2 group (P < 0.0001), and control drug 3 group (P < 0.01), that is, the temperature-lowering effect of the experimental drug was significantly better than that of control drug 1, control drug 2, and control drug 3.

[0081] Table 4 Average body temperature of each group on the 14th day

[0082]

[0083] Note: The code # in the table indicates compared with the model group *** P < 0.0001; the code * respectively indicates compared with the experimental drug group * P < 0.05, ** P < 0.01, ****P<0.0001.

[0084] 8.3 Lung and thymus organ index

[0085] The lung and thymus tissue organ indexes of pigs were analyzed. Figure 2 shown.

[0086] Compared with the model group, the drug-treated groups (P<0.0001) were able to significantly reduce the virus-induced pulmonary enlargement organ lesions, which was statistically significant; compared with the experimental drugs, control drug 1 (P<0.05), control drug 2 (P<0.01), and control drug 3 (P<0.01) had statistical differences.

[0087] The experimental drug group was able to alleviate the thymic atrophic organ lesions caused by the virus, which was statistically significant (P<0.01). The control drug groups 1, 2 and 3 only showed a trend of alleviating lung organ enlargement and thymic atrophy, and did not show a significant effect. Compared with the experimental drug, control drug 1 (P<0.05), control drug 2 (P<0.05) and control drug 3 (P<0.01) were statistically different.

[0088] That is, the test drug is superior to control drug 1, control drug 2 and control drug 3 in reducing lung organ enlargement and thymus atrophy.

[0089] 8.4 Histopathological examination

[0090] The lung pathological sections of the pigs in each group were as follows: Figure 3 As shown. The pigs in the blank group had normal lungs and no pathological damage was found; the pigs in the model group had obvious interstitial pneumonia, thickened alveolar walls, lymphocyte infiltration, cavitary hemorrhage, and vascular hemorrhage; compared with the model group, the experimental drug group, control drug group 1, control drug group 2, and control drug group 3 improved lung damage to varying degrees, reducing the degree of alveolar wall thickening, bleeding, and lymphocyte infiltration. The results showed that drug treatment can reduce lung tissue damage and lesions caused by blue ear disease virus infection, and the experimental drug is better than control drug 1, control drug 2, and control drug 3 in improving histopathological damage.

[0091] 8.5 Lung Viral Load Detection

[0092] Lung viral load results Figure 4As shown. No virus was detected in the blank group; compared with the model group, the test drug group detected a lower viral load than the model group, which was statistically significant (P<0.0001); control drug 1, control drug 2, and control drug 3 had a trend of reducing the amount of virus, and there was no statistical difference. Compared with the test drug group, control drug 1 (P<0.05), control drug 2 (P<0.05), and control drug 3 (P<0.01) were statistically different. The results showed that the drug combination of the test drug was superior to control drug 1, control drug 2, and control drug 3, and could significantly inhibit the effect of blue ear disease virus in the lungs of sick pigs.

[0093] 9. Conclusion

[0094] In summary, the results show that the Huashituidu granules involved in the present invention have the effect of treating blue ear disease. At the macro level, it can improve the respiratory symptoms of pigs and lower body temperature; at the micro level, it can reduce organ lesions of lung enlargement and thymus atrophy, improve histopathological damage, and reduce the role of lung viral load. Comprehensive comparison shows that the effect of the experimental drug in treating blue ear disease in pigs is better than that of control drug 1 (the original Huashituidu prescription), control drug 2 (Qingwen Baidusan), and control drug 3 (CN 116077578 A Example 3 formula drug). In particular, the effect of the experimental drug is better than that of control drug 1 (the original Huashituidu prescription), indicating that the application reuses patchouli and gypsum as the monarch, increases the dosage of the ministerial drug Magnolia officinalis, changes the treatment principle of the original prescription, and the technical effect achieved is unexpected.

[0095] The Hua Shi Bai Du granules involved in the present invention can improve the symptoms of blue ear disease caused by porcine reproductive and respiratory syndrome virus, reduce lung lesions and damage at the macroscopic and microscopic levels, significantly reduce lung viral load, reduce lung enlargement and thymus atrophy, have the effect of treating blue ear disease, and have good prospects for application in this aspect.

Claims

1. Application of Huashibaidu granules in the preparation of drugs for treating blue ear disease in pigs; wherein: The effective ingredients contained in every 1000g of the Huashi Baidu granules are made of the following raw materials in percentage by weight: 14-20% of patchouli, 14-20% of gypsum, 7-10% of magnolia bark, 7-10% of atractylodes, 7-10% of poria, 4-6% of pinellia, 3-4% of ephedra, 4-6% of stir-fried bitter almonds, 5-7% of tsaoko, 2-3% of raw rhubarb, 5-7% of red peony root, 5-7% of scutellaria baicalensis, 5-7% of astragalus, and 1-2% of licorice.

2. The use according to claim 1, characterized in that: The effective ingredients contained in every 1000g of the Hua Shi Baidu granules are made of the following raw materials in percentage by weight: 17.0% patchouli, 17.0% gypsum, 8.5% magnolia bark, 8.5% atractylodes, 8.5% tuckahoe, 5.1% pinellia, 3.4% ephedra, 5.1% stir-fried bitter almonds, 5.6% tsaoko, 2.8% raw rhubarb, 5.6% red peony root, 5.6% scutellaria seed, 5.6% astragalus, and 1.7% licorice.

3. The use according to claim 1 or 2, characterized in that: The effective component is prepared by the following method: add gypsum to 10 times of water and boil for 20 minutes, then add ephedra, fried bitter almond, licorice, magnolia bark, atractylodes, tsaoko, pinellia tuber, tuckahoe, astragalus, scutellaria baicalensis and red peony root and boil for 30 minutes, then add patchouli and rhubarb and boil for 10 minutes, filter to obtain the first decoction; add 8 times of water to the residue and boil for 40 minutes, filter to obtain the second decoction; combine the two decoctions, and concentrate under reduced pressure at 65°C to obtain an extract with a relative density of 1.06-1.20 at 25°C, so as to obtain the effective component.

Images