Traditional Chinese medicine gel for treating thyroid nodules from liver as well as preparation method and application of traditional Chinese medicine gel

Abstract

The invention relates to traditional Chinese medicine gel for treating thyroid nodules from liver as well as a preparation method and application of the traditional Chinese medicine gel. The traditional Chinese medicine gel takes an extracting solution of a traditional Chinese medicine composition as an effective component in parts by weight, the traditional Chinese medicine composition comprises the following raw materials: 15-25 parts of selfheal, 10-20 parts of radix bupleuri, 8-15 parts of vinegar-processed curcuma zedoary, 8-15 parts of fructus forsythiae, 5-15 parts of thunberg fritillary bulb, 5-15 parts of moutan bark, 5-8 parts of scutellaria baicalensis, 5-10 parts of oyster, 3-8 parts of mint and 3-8 parts of borneol. From the perspective of soothing liver-qi stagnation, the thyroid nodules are treated by firstly soothing liver-qi stagnation and cooperating with phlegm reducing, stagnation resolving, blood circulation promoting and blood stasis removing, mint and borneol are added in the formula, and the overall improvement effect of the external medicine on unhealthy emotions such as depression and anxiety and the dissipation effect on the nodules are enhanced. Meanwhile, all the medicinal materials in the formula are classified, extracted and refined, all the effective components are reserved to the maximum extent, transdermal absorption is good, and the effects of reducing swelling, removing stasis and improving the depressive emotion are improved.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine gel for treating thyroid nodules from the liver, and a preparation method and application thereof. Background Art

[0002] Thyroid nodules refer to discrete lesions caused by abnormal, focal growth of thyroid cells in the thyroid gland, which fall into the category of "goiter" and "goiter" in traditional Chinese medicine. With the improvement of medical conditions and the improvement of various imaging tests, the detection rate of thyroid nodules is also increasing. The 2023 "Guidelines for the Diagnosis and Treatment of Thyroid Nodules and Differentiated Thyroid Cancer (Second Edition)" points out that the prevalence of thyroid nodules in adults is as high as 20.43%. The high incidence of thyroid nodules brings huge economic and psychological burdens to people, seriously endangering people's quality of life.

[0003] At present, surgical treatment, radioactive iodine treatment, anhydrous ethanol injection treatment, ultrasound-guided ablation treatment and thyroid hormone suppression treatment are mostly used in clinical treatment, but these methods have certain disadvantages such as adverse reactions and side effects, high treatment risks, limited scope of application, and high costs. Traditional Chinese medicine starts from the overall perspective, treats both the symptoms and the root causes, has flexible methods, good efficacy, and few adverse reactions, and has become a better choice for the treatment of thyroid nodules. Traditional Chinese medicine believes that qi stagnation, phlegm coagulation, and blood stasis are the basic pathological changes of thyroid nodules. Relevant scholars have proposed the theory that "phlegm and blood stasis" cause goiter, and pay more attention to the interaction between phlegm and blood stasis and the changes in the process of goiter. Therefore, more attention is paid to the principle of phlegm and blood stasis, and soft nodules in diagnosis and treatment. However, due to fierce competition in modern society, depression, anxiety and other negative emotions caused by long-term life and work pressure are currently the main risk factors for the occurrence of thyroid nodules. Depression, anxiety and other negative emotions can cause liver disorders, abnormal drainage, and liver qi stagnation, which in turn causes qi stagnation, phlegm coagulation, and blood stasis in the neck, resulting in nodules. The key to the pathogenesis is that the liver qi stagnates first, which in turn produces phlegm and turns into blood stasis.

[0004] In addition, there are some other problems in the field of TCM treatment of thyroid nodules, such as the lack of research on the external treatment of thyroid nodules. As a commonly used external treatment method, the application of Chinese medicine external preparations has unique advantages in the treatment of superficial diseases such as thyroid nodules. It can reduce the stimulation of the gastrointestinal tract caused by long-term oral administration and the toxic and side effects on organs such as the liver and kidneys. It has a wide range of applications and is easier to be accepted by patients. However, there are relatively few Chinese medicine external preparations that can be used to treat thyroid nodules. Most of the existing external preparations are made by grinding Chinese medicine into fine powder and adding vaseline, ginger juice, wine, honey, etc., which have the disadvantages of low transdermal absorption efficiency, inconvenient use, easy to fall off, and affecting the appearance. In summary, there is an urgent need for a Chinese medicine external preparation that can treat thyroid nodules from the liver and is convenient, beautiful, safe and effective in clinical practice to meet the huge social and medical needs. Summary of the invention

[0005] In order to overcome the problems existing in the prior art, the purpose of the present invention is to provide a traditional Chinese medicine gel for treating thyroid nodules from the liver, and a preparation method and application thereof.

[0006] In order to achieve the above-mentioned purpose and other related purposes, the present invention adopts the following technical solutions:

[0007] The first aspect of the present invention is to provide a Chinese medicine composition for treating thyroid nodules by treating the liver. The composition treats thyroid nodules by soothing the liver and relieving depression first, and cooperates with the methods of resolving phlegm and dispersing nodules and promoting blood circulation and removing blood stasis.

[0008] The embodiment of the present application provides a traditional Chinese medicine composition for treating thyroid nodules by treating the liver, which comprises the following raw materials in parts by weight: 15-25 parts of Prunella Vulgaris, 10-20 parts of Bupleurum chinense, 8-15 parts of Curcuma zedoariae, 8-15 parts of Forsythia suspensa, 5-15 parts of Fritillaria thunbergii, 5-15 parts of Paeonia suffruticosa, 5-8 parts of Scutellaria baicalensis, 5-10 parts of oyster, 3-8 parts of mint and 3-8 parts of borneol.

[0009] The inventor of the present application attaches importance to the important role played by negative emotions such as depression and anxiety in the formation of the disease at this stage when diagnosing and treating thyroid nodules. It is believed that negative emotions such as depression and anxiety can cause liver qi stagnation and lead to liver qi stagnation. Liver qi stagnation occupies an important position in the onset of thyroid nodules and is the basic pathogenesis throughout thyroid nodules. Therefore, the method of treating from the liver is adopted, starting from the perspective of soothing the liver and relieving depression, taking soothing the liver and relieving depression as the first, and cooperating with the method of resolving phlegm and dispersing nodules, promoting blood circulation and removing blood stasis to treat thyroid nodules. The main medicines in the prescription are Prunella vulgaris and Bupleurum. Prunella vulgaris belongs to the liver and gallbladder meridians, reduces swelling and dissipates nodules, and is mainly used to treat goiters. The "Shennong Bencao Jing" says that it "mainly treats cold and heat, scrofula... eliminates goiter and qi stagnation", which shows that Prunella vulgaris can disperse depression and eliminate goiters. The Illustrated Classic of Materia Medica records that it "enters the Jueyin meridian alone", "replenishes the Jueyin blood vessels, and can also dredge qi stagnation". The Southern Yunnan Materia Medica states that it "enters the liver meridian... relieves liver depression and promotes liver qi". This shows that Prunella vulgaris has certain effects in regulating liver qi and relieving liver depression. Bupleurum belongs to the liver and gallbladder meridians, can remove qi accumulation, dredge liver wood, and bring forth the new. The Righteousness of Materia Medica states that Bupleurum "relaxes the qi of the liver and gallbladder", so Bupleurum is particularly good at treating qi stagnation in the liver and gallbladder. The Explanation of the Classic of Materia Medica records that "Bupleurum is light and clear... it can disperse any qi stagnation". The assistant drugs are vinegar-cured zedoaria, forsythia, Fritillaria thunbergii, and peony bark. Vinegar-cured zedoaria belongs to the liver and spleen meridians, promotes qi and blood circulation, and removes blood stasis. The Medical Heart Method states that "Guangshen is zedoaria. It is used for promoting qi and blood circulation, removing accumulation and dispersing stagnation". Lianqiao can reduce swelling and dissipate nodules, especially scrofula and phlegm nodules. The Shennong's Herbal Classic records that Lianqiao can treat goiters, scrofula and other diseases. Moutan bark can promote blood circulation and dissipate blood stasis, and it helps the vinegar Curcuma zedoaria to promote blood circulation. The Changsha Yaojie says: "It can promote blood stasis and purge heat, and dissolve the symptoms of the internal organs." Zhejiang Fritillaria can clear heat and resolve phlegm, dissipate nodules and eliminate carbuncle. The Bencao Zheng says that Zhejiang Fritillaria "is the best at reducing phlegm and qi, and is good at relieving depression." The Bencao Fengyuan points out that Zhejiang Fritillaria and Lianqiao are used together to treat tuberculosis on the neck, "both for their ability to dissipate depression and resolve nodules, resolve phlegm and detoxify." The adjuvants are Scutellaria baicalensis and Oyster. Scutellaria baicalensis can reduce swelling and dissipate nodules, and when used with Bupleurum chinense, it can coordinate the rise and fall and make the qi flow smoothly. Oysters soften and disperse nodules. "Compendium of Materia Medica" says: "It can resolve phlegm, soften nodules... and eliminate goiter and tuberculosis"; borneol and mint are both guiding drugs. Mint belongs to the lung and liver meridians, and its fragrance can open the orifices. It has the effects of soothing the liver, relieving depression, and promoting the flow of qi, which helps to improve depression. "Medical Records of Chinese and Western Medicine" says: "Mint is pungent, with a clear and fragrant aroma... It can penetrate the muscles and bones inside and reach the skin surface outside, promote the circulation of internal organs, and run through the meridians"; borneol has the effect of opening all orifices and reducing swelling. "Compendium of Materia Medica" evaluates borneol as "opening all orifices and dispersing depressed fire". The two can be used together to help promote other drugs to reach the diseased area directly, by leveraging its pungent fragrance to reach the skin pores, thereby enhancing the drug's effect on improving depression and dissipating nodules.

[0010] The present invention also provides a traditional Chinese medicine gel for treating thyroid nodules from the liver, which adopts the extract of the traditional Chinese medicine composition; in parts by weight, the raw materials of the traditional Chinese medicine gel are: 40 parts of the extract of the traditional Chinese medicine composition, 1.5 parts of carbomer, 10 parts of glycerol, 0.5 parts of Tween-80, 2 parts of azone and 0.1 parts of ethyl hydroxybenzoate, 100 parts of purified water are added, and triethanolamine is used to adjust the pH to 7.5.

[0011] The preparation method of the traditional Chinese medicine gel comprises the following specific steps:

[0012] (1) weighing the raw materials of the Chinese medicine composition according to proportion and setting aside;

[0013] (2) adding purified water to the mint, slowly heating to boiling, and keeping it at a slight boil for 5 hours, then stopping the heating, leaving it for a while, collecting the volatile oil, and obtaining the mint volatile oil for later use;

[0014] (3) soaking peony bark in pure water, extracting by steam distillation, and collecting distillate; placing the distillate at room temperature, refrigerating for crystallization, filtering by suction, and drying under reduced pressure to obtain drug crystals; mixing the drug crystals with borneol, grinding, adding 95% ethanol to dissolve, and then dissolving peppermint volatile oil therein to obtain solution A;

[0015] (4) adding Forsythia suspensa, Prunella vulgaris, Scutellaria baicalensis, Bupleurum chinense and Oyster to purified water for decoction, concentrating, precipitating with alcohol, standing overnight and centrifuging to obtain a supernatant solution B;

[0016] (5) Add ethanol to extract Curcuma zedoaria and Fritillaria thunbergii, filter, and mix the filtrate to obtain solution C; mix the supernatant solution B and solution C, concentrate, add solution A, and then add 95% ethanol to make up the volume to obtain an extract of the traditional Chinese medicine composition.

[0017] The concentration of the extract of the traditional Chinese medicine composition is 1g of crude drug / mL of the extract of the traditional Chinese medicine composition.

[0018] (6) Weighing the raw materials of the Chinese medicine gel according to proportion;

[0019] (7) After the carbomer is swollen with purified water, triethanolamine is added to adjust the pH to 7.5, and the mixture is aged to obtain a matrix I; the extract of the Chinese medicine composition, glycerol, Tween-80, azone and ethyl hydroxybenzoate are fully mixed and dissolved to obtain a composition II; the matrix I and the composition II are mixed, purified water is added to make up the weight, and the mixture is fully stirred to obtain a Chinese medicine gel.

[0020] In step (4), since the water extract contains macromolecular impurities such as polysaccharides, and the traditional Chinese medicine gel is an external preparation, the macromolecular impurities are not easily absorbed through the skin, affecting the clinical efficacy. At the same time, the large number of macromolecular impurities is not conducive to the formation of the gel in the later stage. Therefore, the present invention removes the macromolecular impurities as much as possible through the alcohol precipitation process on the basis of retaining the effective ingredients.

[0021] In step (6), glycerol is selected and supplemented with Tween-80 as a solvent, which not only improves the solubility of fat-soluble components but also reduces irritation to the skin. Triethanolamine neutralizes with the carboxyl group of carbomer to form a stable polymer structure, thereby achieving the application effects of thickening and moisturizing.

[0022] In step (2), the mass ratio of mint to purified water is 1:8.

[0023] In step (3), the mass ratio of peony bark to purified water is 1:12.

[0024] In step (4), 8 times the amount of purified water is added to the medicinal material by weight, and the mixture is heated and refluxed for extraction 3 times, each time for 1 hour; 95% ethanol is added to the concentrated solution until the final ethanol concentration reaches 70% for alcohol precipitation.

[0025] In step (5), add 6 times the weight of the medicinal material to 60% ethanol, heat and reflux to extract twice, each time for 1.5 hours.

[0026] In step (6), Carbomer 940 is a hydrophilic gel skeleton material, triethanolamine is a pH regulator, glycerol is a moisturizer, Tween-80 is a solubilizer, Azone is a transdermal enhancer, and ethylparaben is a preservative.

[0027] The third aspect of the present invention is to provide an application of the traditional Chinese medicine gel for treating thyroid nodules from the perspective of the liver in basic research and clinical practice.

[0028] Specifically, in basic research, chronic unpredictable mild stress combined with propylthiouracil drug induction was used to prepare a rat model of thyroid nodules with liver qi stagnation. The behavioral evaluation results showed that the model rats showed obvious depression, which was consistent with the symptom characteristics of liver qi stagnation in clinical Chinese medicine; in addition, ultrasound imaging, morphology and organ index determination, and pathological staining results showed that the thyroid glands of the model group rats were significantly enlarged and formed nodular lesions, the thyroid index increased significantly, and the thyroid follicular epithelial cells were highly proliferative, which was consistent with the clinical manifestations and met the evaluation criteria for the rat model of this disease.

[0029] Furthermore, the results of the smearing and drug administration intervention showed that the Chinese medicine gel can significantly improve the depression of rats, significantly reduce the enlargement of the thyroid gland, improve nodular lesions, reduce the thyroid index, and inhibit the abnormal nodular hyperplasia of thyroid follicular epithelial cells. Furthermore, the pharmacological mechanism of this efficacy is that the Chinese medicine gel improves the symptoms of thyroid nodules by increasing free triiodothyronine (FT3) and free thyroxine (FT4) in serum, reducing the level of thyroid stimulating hormone (TSH) and reducing the content of inflammatory factors in thyroid tissue.

[0030] Specifically, in clinical practice, this study selected 58 patients with thyroid nodules who met the criteria and visited the Endocrinology Department of Jinan Central Hospital from December 2022 to December 2023, and used a self-control before-and-after research method to give them the traditional Chinese medicine gel treatment. Apply an appropriate amount of gel to the thyroid skin surface of the neck, and gently massage to promote absorption, 2-3 times a day. After 2 months (60 days) of treatment, the total effective rate of reduction in the maximum diameter of the nodule was 70.69%, the total effective rate of TI-RADS classification changes was 63.79%, and the effective rate of clinical symptom improvement was 82.76%, indicating that the traditional Chinese medicine gel for treating thyroid nodules from the liver can reduce the diameter of the nodules and improve clinical discomfort symptoms.

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] 1. The Chinese medicine compound of the present invention adopts a scientific ratio, and the components are coordinated and complemented with each other to play a synergistic effect. The present invention has obvious innovation and practicality as a prescription for treating thyroid nodules: (1) The prescription is highly unified in terms of prescription and principle. In view of the increasing number of negative emotions such as depression and anxiety in today's society and the increasing incidence of thyroid nodules, it is believed that liver qi stagnation caused by negative emotions such as depression and anxiety plays an important role in the onset of thyroid nodules at this stage. The prescription adopts the method of soothing the liver and relieving depression first, and cooperates with the treatment of resolving phlegm and dispersing nodules, promoting blood circulation and removing blood stasis; (2) Peppermint and borneol are added to the prescription, and their spicy fragrance can reach the skin pores, thereby enhancing the overall improvement of negative emotions such as depression and anxiety and the dissipation of nodules by external drugs. In addition, mint itself has the effects of soothing the liver, relieving depression, and promoting the flow of qi, which helps to enhance the liver-soothing effect and improve negative emotions such as depression and anxiety. (3) The prescription is simple in dosage, inexpensive, and does not contain any rare or precious medicinal materials. It is acceptable to most patients with thyroid nodules. On the basis of good clinical efficacy, it can also reflect a certain degree of economy and is worthy of promotion and application.

[0033] 2. The present invention adopts external treatment to treat thyroid nodules. Compared with oral drugs, the Chinese medicine gel of the present invention can make patients avoid the inconvenience caused by long-term oral drugs, reduce the stimulation to the gastrointestinal tract, and is more easily accepted by patients and has a wider scope of application. Secondly, the present invention is based on the physicochemical properties of active ingredients, and the various medicinal materials in the prescription are classified and extracted by modern technology and the best extraction and refining process are optimized. Compared with the external application of traditional drug grinding fine powder, this retains each effective ingredient to the greatest extent and has good transdermal absorption, which is conducive to improving the effect of treating thyroid nodules. Finally, the gel used in the present invention has suitable fluidity, is easy to spread, has good affinity with the skin, and is convenient and beautiful to use, and does not pollute clothes. Compared with traditional external patches, it has good air permeability, low skin irritation, is not easy to cause skin allergies, and is safer to use.

[0034] 3. The development of TCM drugs for the treatment of thyroid nodules requires a suitable animal model. The animal models currently used for the study of thyroid nodules do not simulate TCM syndromes at the same time, and there is a certain gap with the practice of TCM syndrome differentiation and treatment of thyroid nodules in clinical practice. The present invention provides a rat model of thyroid nodules with liver qi stagnation syndrome, which is consistent with the treatment method of the TCM gel from the liver, and the pharmacodynamic mechanism research conducted on this basis is more scientific and accurate. The TCM gel of the present invention can improve nodular lesions by increasing FT3 and FT4 in rat serum, reducing TSH levels and reducing the content of inflammatory factors in thyroid tissue, and play a role in regulating goiter, abnormal nodular hyperplasia of follicular epithelial cells and depression. In addition, clinical studies have also confirmed that the TCM gel for treating thyroid nodules from the liver can effectively reduce the diameter of nodules and improve clinical discomfort symptoms. The present invention provides a reference and idea for solving the problem of TCM external preparations for the treatment of thyroid nodules that have no efficacy and clear mechanism of action faced by medical technology. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 : Extraction process routes of various medicinal materials in traditional Chinese medicine gel;

[0036] Figure 2 :The molding process of traditional Chinese medicine gel;

[0037] Figure 3 : Comparison of cumulative transdermal curves of paeonol before and after the addition of peppermint oil to traditional Chinese medicine gel;

[0038] Figure 4 : Comparison of the cumulative transdermal curves of baicalin before and after the addition of peppermint oil to traditional Chinese medicine gel;

[0039] Figure 5 : Comparison of the cumulative transdermal curves of forsythiaside A before and after the addition of peppermint oil in traditional Chinese medicine gel;

[0040] Figure 6: Comparison of the cumulative transdermal curves of rosmarinic acid before and after the addition of peppermint oil to traditional Chinese medicine gel;

[0041] Figure 7 : Comparison of behavioral experimental results of rats in each group after drug intervention (n=13); A, B, and C are the comparison of the total movement distance of rats in the open field test, the sugar water preference rate in the sugar water preference test, and the suspension immobility time in the forced swimming test; compared with the blank group, ## P<0.01; compared with the model group, * P<0.05, ** P < 0.01;

[0042] Figure 8 : Comparison of thyroid images of the left and right sides of each group of rats during ultrasonic transverse scanning; A: blank group; B: model group; C: positive drug methyl lemon solution group; D: low-dose group of the Chinese medicine gel of the present invention; E: medium-dose group of the Chinese medicine gel of the present invention; F: high-dose group of the Chinese medicine gel of the present invention; the red arrows mark the images of the left and right sides of the thyroid when the ultrasonic probe is scanned transversely;

[0043] Fig. 9 : Comparison of thyroid images of rats in each group during longitudinal ultrasound scanning; A: blank group; B: model group; C: positive drug methyl lemon solution group; D: low-dose group of the Chinese medicine gel of the present invention; E: medium-dose group of the Chinese medicine gel of the present invention; F: high-dose group of the Chinese medicine gel of the present invention; the red dotted line frame is the image of the thyroid gland during longitudinal ultrasound scanning;

[0044] Fig.10 : Comparison of thyroid volume on the left and right sides of rats in each group (n=10); A and B are the comparison of the left thyroid volume and the right thyroid volume of rats in each group, respectively; compared with the blank group, ## P<0.01; compared with the model group, * P<0.05, ** P < 0.01;

[0045] Fig.11 : Comparison of thyroid morphology of rats in each group; A: blank group; B: model group; C: methyl lemon solution group; D: low-dose group of the Chinese medicine gel of the present invention; E: medium-dose group of the Chinese medicine gel of the present invention; F: high-dose group of the Chinese medicine gel of the present invention;

[0046] Fig.12 : Comparison of thyroid index of rats in each group (n=14); compared with the blank group, ## P<0.01; compared with the model group, * P<0.05, ** P < 0.01;

[0047] Fig.13: Comparison of thyroid tissue pathological examination results of rats in each group (×20); A: blank group; B: model group; C: methyl lemon solution group; D: low-dose group of the Chinese medicine gel of the present invention; E: medium-dose group of the Chinese medicine gel of the present invention; F: high-dose group of the Chinese medicine gel of the present invention; The green arrows mark the thyroid follicles; the red arrows mark the thyroid follicular epithelial cells;

[0048] Fig.14 : Comparison of serum thyroid hormone levels in rats in each group (n=10); A, B, C are comparisons of serum FT3, FT4 and TSH levels in rats in each group, respectively; compared with the blank group, ## P<0.01; compared with the model group, * P<0.05, ** P < 0.01;

[0049] Fig.15 : Comparison of IL-1β, IL-6 and TNF-α protein contents in thyroid tissue of rats in each group (n=8); A, B, C are comparison of IL-1β, IL-6 and TNF-α protein contents in thyroid tissue of rats in each group, respectively; compared with the blank group, ## P<0.01; compared with the model group, * P<0.05, ** P < 0.01;

[0050] Fig.16 : Comparison of thyroid ultrasound examination results of patient 1 before and after treatment; the left examination sheet is the thyroid ultrasound condition before treatment, and the right examination sheet is the thyroid ultrasound condition after treatment. Comparison between the two shows that the diameter of the nodule has been significantly reduced.

[0051] Fig.17 : Comparison of thyroid ultrasound examination results of patient 2 before and after treatment; the left examination sheet shows the TI-RADS 3 grade of the left thyroid nodule before treatment, and the right examination sheet shows the TI-RADS 2 grade of the left thyroid nodule after treatment. It can be seen that the nodule grade is significantly reduced. DETAILED DESCRIPTION

[0052] Example 1: Provide a Chinese medicine gel for treating thyroid nodules by treating the liver

[0053] The raw material composition of the traditional Chinese medicine gel is as follows: 40 parts of the extract of the traditional Chinese medicine composition, 1.5 parts of carbomer, 10 parts of glycerol, 0.5 parts of Tween-80, 2 parts of azone and 0.1 parts of ethyl hydroxybenzoate, supplemented with 100 parts of purified water, and triethanolamine is used to adjust the pH to 7.5.

[0054] The Chinese medicine composition comprises the following raw materials by weight: 25 parts of Prunella Vulgaris, 20 parts of Bupleurum Root, 8 parts of Curcuma Rhizoma, 8 parts of Forsythia suspensa, 15 parts of Fritillaria thunbergii, 5 parts of Paeonia suffruticosa, 6 parts of Scutellaria baicalensis, 10 parts of Oyster, 8 parts of Mint and 5 parts of Borneol.

[0055] The specific preparation steps of Chinese medicine gel are as follows:

[0056] (1) weighing the raw materials of the Chinese medicine composition according to proportion and setting aside;

[0057] (2) Add 8 times the amount of purified water and a number of glass beads to the mint by weight, shake and mix, connect the volatile oil analyzer and the reflux condenser, place in an electric heating mantle and slowly heat to boiling, and keep it at a slight boiling for 5 hours, then stop heating, let it stand for a while, collect the volatile oil and set aside;

[0058] (3) Add 12 times the amount of pure water to the peony bark medicinal material by weight and soak for 30 minutes, extract it by steam distillation, and collect about 6 times the amount of distillate. The distillate is placed at room temperature for 1 hour, refrigerated for 12 hours (overnight) to precipitate crystals, filtered, and dried at 45°C under reduced pressure for 1.5 hours to obtain drug crystals. The drug crystals are mixed with borneol by weight, fully ground, and a small amount of 95% ethanol is added to dissolve, and then peppermint volatile oil is dissolved therein to obtain solution A;

[0059] (4) Add weight parts of Forsythia suspensa, Prunella vulgaris, Scutellaria baicalensis, Bupleurum chinense and Oyster to pure water for decoction, and then concentrate the decoction to 1 g crude drug / ml. After the concentrate is cooled to room temperature, ethanol is added for alcohol precipitation, and the concentrate is allowed to stand at 4°C overnight, and centrifuged to obtain a supernatant solution B;

[0060] (5) Add ethanol to extract the weight parts of Curcuma zedoaria and Fritillaria thunbergii. Then filter and mix the filtrate to obtain solution C. Mix solutions B and C, add solution A after concentration, and then add 95% ethanol to make up the volume, and finally obtain an extract of the Chinese medicine composition with a concentration of 1 g crude drug / mL;

[0061] (6) Weighing the raw materials of the Chinese medicine gel according to proportion;

[0062] (7) Carbomer 940 was swollen with 15 times the amount of purified water for 12 hours, triethanolamine was added to adjust the pH to 7.5, and the mixture was further aged for 12 hours to obtain matrix I. The extract of the Chinese medicine composition, glycerol, Tween-80, azone and ethyl hydroxybenzoate were fully mixed and dissolved to obtain composition II. Matrix I and composition II were mixed, purified water was added to make up the weight, and the mixture was fully stirred to obtain a Chinese medicine gel.

[0063] Example 2: Provide a Chinese medicine composition for treating thyroid nodules by treating the liver

[0064] The raw material composition of the traditional Chinese medicine gel is as follows: 40 parts of the extract of the traditional Chinese medicine composition, 1.5 parts of carbomer, 10 parts of glycerol, 0.5 parts of Tween-80, 2 parts of azone and 0.1 parts of ethyl hydroxybenzoate, supplemented with 100 parts of purified water, and triethanolamine is used to adjust the pH to 7.5.

[0065] The Chinese medicine composition comprises the following raw materials by weight: 15 parts of Prunella Vulgaris, 15 parts of Bupleurum Root, 10 parts of Curcuma Rhizoma, 10 parts of Forsythia suspensa, 5 parts of Fritillaria thunbergii, 13 parts of Paeonia suffruticosa, 8 parts of Scutellaria baicalensis, 5 parts of Oyster, 3 parts of Menthus Fructus and 8 parts of Borneolum Syntheticum.

[0066] The specific preparation steps of the Chinese medicine gel are the same as those in Example 1.

[0067] Example 3: Provide a Chinese medicine composition for treating thyroid nodules by treating the liver

[0068] The raw material composition of the traditional Chinese medicine gel is as follows: 40 parts of the extract of the traditional Chinese medicine composition, 1.5 parts of carbomer, 10 parts of glycerol, 0.5 parts of Tween-80, 2 parts of azone and 0.1 parts of ethyl hydroxybenzoate, supplemented with 100 parts of purified water, and triethanolamine is used to adjust the pH to 7.5.

[0069] The Chinese medicine composition comprises the following raw materials by weight: 20 parts of Prunella Vulgaris, 10 parts of Bupleurum Root, 9 parts of Curcuma Rhizoma, 15 parts of Forsythia suspensa, 10 parts of Fritillaria thunbergii, 10 parts of Paeonia suffruticosa, 5 parts of Scutellaria baicalensis, 5 parts of Oyster, 8 parts of Menthus fragrans and 3 parts of Borneolum.

[0070] The specific preparation steps of the Chinese medicine gel are the same as those in Example 1.

[0071] Example 4: Preparation technology and transdermal absorption study of Chinese medicine gel for treating thyroid nodules from the perspective of liver

[0072] 1 Study on gel preparation process

[0073] 1.1 Experimental Materials

[0074] 1.1.1 Experimental instruments

[0075]

[0076]

[0077] 1.1.2 Experimental reagents

[0078]

[0079] 1.2 Experimental methods

[0080] 1.2.1 Extraction process route

[0081] The extraction process of each medicinal material of the traditional Chinese medicine gel for treating thyroid nodules from the liver is shown in Figure 1 .

[0082] 1.2.2 Study on the extraction process of peony bark and mint

[0083] The team has optimized and determined the best extraction process for peony bark and mint in their previous work on the "National Major New Drug Creation Project".

[0084] The best extraction process of peony bark: soak the peony bark in 12 times the amount of pure water for 30 minutes, extract it by steam distillation, and collect about 6 times the amount of distillate. The distillate is placed at room temperature for 1 hour, refrigerated for crystallization for 12 hours (overnight), filtered, and dried at 45℃ under reduced pressure for 1.5 hours to obtain drug crystals.

[0085] The best extraction process for mint: add 8 times the amount of pure water and several glass beads to mint, shake to mix, connect the volatile oil analyzer and the reflux condenser, place it in an electric heating jacket and slowly heat to boiling, and keep it at a slight boiling for 5 hours, then stop heating, let it stand for a while, and collect the volatile oil.

[0086] 1.2.3 Study on the water extraction and alcohol precipitation process of Forsythia suspensa and Prunella vulgaris

[0087] 1.2.3.1 Orthogonal optimization study of water extraction process

[0088] Weigh 9 portions of Forsythia suspensa, Prunella vulgaris, Scutellaria baicalensis, Bupleurum chinense and Oyster slices in the prescription ratio, add appropriate amount of water and extract according to the extraction conditions screened by orthogonal design. The amount of water added, extraction time and number of times are the main factors affecting the water extraction efficiency. The contents of forsythiaside A, rosmarinic acid and extracts are used as indicators to investigate the above three factors. The weight coefficients of forsythiaside A, rosmarinic acid and extracts are set as 0.4, 0.4 and 0.2, respectively, as shown in Table 1.

[0089] Table 1. Level table of factors in orthogonal experiment of water extraction process

[0090]

[0091] 1.2.3.2 Alcohol precipitation process research

[0092] The experiment investigated the removal effect of 50%, 70% and 90% ethanol, and used forsythiaside A, rosmarinic acid and extract content as indicators. Weigh the prescription ratio of Forsythia, Prunella Vulgaris, Scutellaria, Bupleurum, and Oyster slices, extract according to the best water extraction process, concentrate to 1g crude drug / ml, divide into 3 parts, add ethanol to make the alcohol concentration reach 50%, 70%, and 90%, stand overnight, centrifuge to obtain the supernatant, and determine the content of forsythiaside A, rosmarinic acid and extract.

[0093] 1.2.4 Study on the alcohol extraction process of vinegar-curdled zedoaria and Fritillaria thunbergii slices

[0094] Weigh 9 portions of vinegar-curd zedoaria and thunbergii bulb slices in the prescription ratio, add an appropriate amount of ethanol of a certain concentration, and then extract under the extraction conditions screened by orthogonal design. The ethanol amount, ethanol concentration, extraction time and number of extractions are the main factors affecting the ethanol extraction efficiency. The contents of germacron and extract are used as indicators to investigate the four factors, and the weight coefficients of the contents of germacron and extract are set as 0.7 and 0.3, respectively, as shown in Table 2.

[0095] Table 2 Alcohol extraction process orthogonal experimental factor level table

[0096]

[0097] 1.2.5 Molding process research

[0098] 1.2.5.1 Molding process route

[0099] The molding process of the Chinese medicine gel for treating thyroid nodules from the liver is shown in Figure 2 .

[0100] 1.2.5.2 Screening of matrix and additives

[0101] Carbomer-940 is selected as the matrix of the gel, and the concentration is between 1% and 2%; triethanolamine is selected as the pH regulator, and the pH is adjusted to 7.5, which can ensure the stability and adhesion of the drug and reduce the irritation of the preparation to the skin; glycerol is not only important for the stability of the dosage form but also conducive to maintaining the moist environment of the skin and has a good moisturizing effect, so glycerol is selected as the moisturizer and the amount of glycerol is determined to be between 5% and 15%; Tween-80 is used as a solubilizer to improve the solubility of the fat-soluble extract in the water-soluble matrix, and the amount of Tween-80 is between 0.5% and 1.5%; 0.1% of ethyl hydroxybenzoate is selected as a preservative; azone usually has the best penetration-promoting effect when the mass fraction is 2%.

[0102] 1.2.5.3 Orthogonal experimental optimization of molding process

[0103] On the basis of the preliminary experiment, 40g of the extract of the Chinese medicine composition was fixed, so that the mass of the auxiliary material was 60g. The effects of the addition amount of gel matrix Carbomer 940, moisturizer glycerol, and solubilizer Tween-80 on the uniformity, viscosity, and stability of the gel were investigated. This study scored each investigation index according to Table 3, with a weighted coefficient of 0.2, and finally a comprehensive score was performed. The concentration of Carbomer-940, the dosage of Tween-80 and glycerol were investigated by orthogonal experiment, and the optimal parameters were determined. The level of the formula ratio factor is shown in Table 4.

[0104] Table 3 Gel inspection index scoring table

[0105]

[0106]

[0107] Table 4. Formula ratio factor level table

[0108]

[0109] 1.3 Experimental Results

[0110] 1.3.1 Orthogonal optimization study of water extraction process

[0111] The orthogonal experimental results of water extraction process are shown in Table 5, and the variance analysis results are shown in Table 6.

[0112] Table 5 Water extraction process orthogonal test results

[0113]

[0114] Table 6 Analysis of variance

[0115]

[0116]

[0117] There are significant differences between the levels of the extraction times influencing factor, and C3 is the best, that is, the best extraction times are 3 times; there is no significant difference between the levels of the extraction time influencing factor, and the intuitive analysis results show that the difference between the levels is small. Considering the cost and time in the production process, A1 is selected; similarly, the intuitive analysis results show that B1 is significantly different from B2 and B3. Considering the actual situation such as cost, B2 is selected. Therefore, the best combination is A1B2C3, that is, the best water extraction process is: 8 times the amount of water, decocted 3 times, each time for 1 hour.

[0118] 1.3.2 Alcohol precipitation process research

[0119] The results of the alcohol precipitation process test are shown in Table 7.

[0120] Table 7 Comparison of alcohol precipitation effects at different ethanol concentrations

[0121]

[0122] The effect of alcohol precipitation with 50% ethanol is poor, while that with 70% and 90% ethanol is good, but the loss of effective ingredients is large and the amount of ethanol used is also high. This study starts from the perspective of retaining effective ingredients to the maximum extent while removing impurities, while taking into account the reduction of production costs, and finally selects 70% ethanol concentration for alcohol precipitation.

[0123] 1.3.3 Orthogonal optimization study of alcohol extraction process

[0124] The orthogonal experimental results of alcohol extraction process are shown in Table 8, and the variance analysis results are shown in Table 9.

[0125] Table 8 Orthogonal experimental results of alcohol extraction process

[0126]

[0127]

[0128] Table 9 Analysis of variance

[0129]

[0130] Among the influencing factors, there was no significant difference between the levels of the extraction times influencing factor, but the intuitive analysis results showed that D1 had a large difference compared with D2 and D3. Considering the actual situation such as cost and safety, D2 was selected, that is, the extraction times were 2 times, which was the best; there was no significant difference between the levels of the extraction time influencing factor, but the intuitive analysis results showed that C1 had a large difference compared with C2 and C3, and the difference between C2 and C3 was small. Considering the cost and time in the production process, C2 was selected; there was no significant difference between the levels of the ethanol addition and ethanol concentration influencing factors, and A1B1 was the best, that is, 6 times the amount of 60% ethanol was used for decoction. Therefore, the best combination is A1B1C2D2, that is, the best alcohol extraction process is: 6 times the amount of 60% ethanol, decoction 2 times, each time 1.5h.

[0131] 1.3.4 Orthogonal optimization study of molding process

[0132] The orthogonal experimental results of gel forming process are shown in Table 10, and the variance analysis results are shown in Table 11.

[0133] Table 10 Orthogonal experimental results of gel forming process

[0134]

[0135]

[0136] Table 11 Analysis of variance

[0137]

[0138] There were significant differences among the levels of factor A (P<0.05), with A2 being the best, i.e., the carbomer concentration was 1.5%. There were no significant differences among the levels of factors B and C. The optimal process parameters were A2B2C1, i.e., the concentration of carbomer-940 was 1.5%, and the addition amounts of glycerol and Tween-80 were 10% and 0.5%, respectively.

[0139] 2 In vitro transdermal absorption study of gel

[0140] 2.1 Experimental Materials

[0141] 2.1.1 Experimental instruments

[0142]

[0143] 2.1.2 Experimental reagents

[0144] The experimental reference substances of paeonol, forsythiaside A, rosmarinic acid and baicalin were provided by Chengdu Munster Biotechnology Co., Ltd.

[0145] 2.2 Experimental methods

[0146] 2.2.1 Selection of the best receiving solution

[0147] In this experiment, 30% and 50% anhydrous ethanol were added to physiological saline, respectively, and the cumulative release amounts of paeonol, forsythiaside A, baicalin and rosmarinic acid in the gel in the above receiving solutions were determined to screen the best receiving solution.

[0148] 2.2.1.1 Cumulative release determination

[0149] The experiment was conducted using a transdermal absorption instrument, with a receiving chamber diameter of 1.4 cm and a receiving pool volume of 15 mL. The receiving chamber was filled with receiving solution (normal saline, normal saline containing 30% ethanol, normal saline containing 50% ethanol), and a 0.45 μm microporous filter membrane was cut into corresponding sizes and tightly attached to the receiving chamber, and bubbles were removed after contact with the receiving solution. 1.4 g of gel was taken and evenly added to the supply chamber, maintained in a constant temperature water bath at 37 ± 0.5 ° C, with the speed set at 300 r / min, and 6 mL of samples were taken at 1, 2, 4, 6, 8, 12, and 24 h after administration, and an equal amount of blank receiving solution at the same temperature was added.

[0150] 2.2.1.2 Determination of forsythiaside A content

[0151] Chromatographic conditions and gradient elution program:

[0152] Table 12 Chromatographic conditions for determination of forsythiaside A content

[0153]

[0154] Table 13 Forsythiaside A gradient elution program

[0155]

[0156] Preparation of test solution: 5 ml of receiving solution was evaporated to dryness at 40 °C and the residue was dissolved in 1 mL of methanol.

[0157] Preparation of reference solution: Take an appropriate amount of Forsythiaside A reference substance and add 50% methanol containing 0.5% glacial acetic acid to make the reference substance solution at 0.0543 mg / mL.

[0158] 2.2.1.3 Determination of rosmarinic acid and baicalin content

[0159] Chromatographic conditions and gradient elution program:

[0160] Table 14 Chromatographic conditions for determination of rosmarinic acid and baicalin content

[0161]

[0162]

[0163] Table 15 Gradient elution program of rosmarinic acid and baicalin

[0164]

[0165] Preparation of test solution: Use the test solution under “2.2.1.2 Determination of Forsythiaside A Content”.

[0166] Preparation of reference solution: Take an appropriate amount of rosmarinic acid reference substance, add 50% ethanol to make up to 0.0459 mg / mL reference substance solution; take an appropriate amount of baicalin reference substance, add methanol to make up to 0.1263 mg / mL reference substance solution.

[0167] 2.2.1.4 Determination of paeonol content

[0168] Chromatographic conditions and gradient elution program:

[0169] Table 16 Chromatographic conditions for determination of paeonol content

[0170]

[0171] Table 17 Paeonol gradient elution program

[0172]

[0173] Preparation of test solution: Accurately pipette 1 ml of receiving solution and filter it through a 0.22 μm microporous filter membrane.

[0174] Preparation of reference solution: Take an appropriate amount of paeonol reference substance and add methanol to make up to 0.0210 mg / mL reference substance solution.

[0175] 2.2.1.5 Data processing

[0176] The following formula is used to calculate the cumulative release of the index component.

[0177]

[0178] Where: Q nis the cumulative release amount per unit area (μg / cm 2 ) of a certain component for the nth sampling, C n is the concentration (μg / mL) of a certain component in the receiving solution for the nth sampling, C i is the mass concentration (μg / mL) of a certain component in the receiving solution for the ith (i < n - 1) sampling, V is the volume of the receiving chamber (15 mL), V0 is the sampling volume (6 mL), and S is the effective permeation area (cm 2 ) during drug diffusion.

[0179] 2.2.2 In vitro transdermal experiment

[0180] The experiment was carried out using the best receiving solution screened by the above research. The rat skin was cut into appropriate sizes and closely attached to the receiving chamber. The remaining steps were the same as those in the method described in "2.2.1.1" of this part. The content determination methods of forsythoside A, rosmarinic acid, baicalin, and paeonol were the same as those in the above research.

[0181] 2.2.3 Determination of skin retention amount

[0182] After the 24-hour transdermal experiment, the skin was taken out, washed clean with physiological saline, the effectively permeated skin was cut out and shredded, 1.0 mL of methanol was added, ultrasonicated for 30 min, and the supernatant was taken by centrifugation.

[0183] 2.3 Experimental results

[0184] 2.3.1 Selection of the best receiving solution

[0185] Table 18 Cumulative release amounts of paeonol, forsythoside A, baicalin, and rosmarinic acid in different receiving solutions

[0186]

[0187]

[0188] The results in Table 18 show that the cumulative release amounts of each component measured in physiological saline are relatively small, and the higher the ethanol content, the higher the cumulative release amounts of each component. Some studies have pointed out that when the ethanol content is relatively high, the skin folds unfold, the hair follicle openings dilate, and the normal physiological structure of the skin changes. Therefore, in this experiment, 30% ethanol - physiological saline was finally used as the receiving solution.

[0189] 2.3.2 Determination of cumulative transdermal amount

[0190] Table 19 Cumulative transdermal amounts of each component in the gel

[0191]

[0192] From Table 19, Figure 3-6The results of in vitro permeation test showed that after adding peppermint oil, the cumulative permeation amount per unit area of ​​paeonol, forsythiaside A, baicalin and rosmarinic acid was 270.54 μg / cm2 respectively within 24 hours. 2 , 3.27μg / cm 2 , 22.30μg / cm 2 , 9.11μg / cm 2 Without peppermint oil, the cumulative permeation per unit area of ​​paeonol, forsythiaside A, baicalin and rosmarinic acid was 250.21 μg / cm2 in 24 hours. 2 , 2.87μg / cm 2 , 20.44μg / cm 2 , 8.79μg / cm 2 , it can be seen that the cumulative amount of active ingredients permeated through the skin is significantly increased after adding peppermint oil.

[0193] 2.3.3 Determination of skin retention

[0194] Table 20 Skin retention of each component in the gel

[0195]

[0196] The experimental results in Table 20 show that the 24h skin retention per unit area of ​​paeonol, forsythiaside A, baicalin and rosmarinic acid are 3.41 μg / cm 2 , 0.18μg / cm 2 , 2.24μg / cm 2 , 0.43μg / cm 2 Among the four ingredients, paeonol had the highest 24-hour skin retention per unit area.

[0197] Example 5: Study on the efficacy and pharmacological mechanism of Chinese medicine gel for treating thyroid nodules from the perspective of liver in rats with thyroid nodules and liver qi stagnation syndrome

[0198] 1 Experimental Materials

[0199] 1.1 Experimental animals

[0200] 84 SPF Wistar rats, weighing 180-220 g, 6-8 weeks old, half male and half female, were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. [SCXK (Beijing) 2021-0011]. The rats were placed in an SPF laboratory with free access to water and food, and the experimental operation was performed after 7 days of adaptive feeding. The experimental plan was approved by the Welfare Ethics Review Committee of the Experimental Animal Center of Jinan Central Hospital.

[0201] 1.2 Experimental instruments

[0202]

[0203] 1.3 Experimental reagents

[0204]

[0205] 2 Experimental methods

[0206] 2.1 Animal grouping, modeling, medication, and sample collection

[0207] After one week of adaptive feeding for all rats, they were evenly divided into 6 groups according to their body weight and the comprehensive scores of various behavioral experiments, namely, a blank group, a model group, a low-, medium- and high-dose group of the Chinese medicine gel of the present invention, and a positive drug methyl lemon solution external preparation group, with 14 rats in each group, 7 males and 7 females. The experiment adopted the method of drug administration while modeling. The rats in the blank group were group-raised, and the rats in the other groups were raised in single cages and the experiment was carried out according to the set CUMS modeling program (Table 21), and the process lasted for 6 weeks. In addition, except for the blank group rats gavaged with an equal volume of normal saline, the rats in the other groups were gavaged with a PTU solution with a concentration of 0.1% at a dose of 1 mL / 100 g once a day, and each group was gavaged for 6 consecutive weeks. In terms of drug administration, the blank group and model group rats were smeared with blank matrix at a dose of 0.52g / kg per day, the low, medium and high dose groups of the Chinese medicine gel of the present invention were smeared with gel at a dose of 0.26, 0.52, and 1.04g / kg per day, respectively, and the positive drug methyl lemon solution external preparation group rats were smeared with methyl lemon solution (E Suizhou Medical Preparation No. 20190014) at a dose of 0.52g / kg per day, and the drug was continuously administered for 6 weeks. After the modeling and drug administration were completed at the same time, behavioral evaluation was performed and thyroid ultrasound examination was performed. After the test, all rats were fasted but not watered for 12h, and then the rats were weighed and anesthetized for breathing, and the rats were fixed for blood sampling from the abdominal aorta, and the upper serum was collected and placed in a -80℃ refrigerator for standby use. The thyroid tissue of the rats after blood sampling was quickly and accurately removed, the thyroid tissue was photographed (with trachea), and then the trachea was removed, the left and right thyroid glands were separated, weighed and recorded respectively, and then a part of the tissue was fixed in 4% paraformaldehyde, and the remaining tissue was placed in a -80℃ refrigerator for standby use.

[0208] Table 21CUMS modeling schedule

[0209]

[0210] Note: Fasting: No feed for 24 hours; Water deprivation: No drinking water for 24 hours; Noise disturbance: Play noise audio at a volume of 110 decibels for 2 hours; Humid environment: Add water to dry bedding to maintain humidity at 60%-70% for 24 hours; Crowded living: Put rats of the same sex in cages, 6 rats per cage, for 24 hours; Restraint stimulation: Put rats in restraint barrels and restrain them for 3 hours; Empty bottle stimulation: Place an empty drinking bottle for 1 hour after 24 hours of water deprivation; Restricted food intake: Give 2 grains of feed after 24 hours of fasting. Feed for 1 hour; cage tilt: raise one end of the cage with feeding trough and water bottle so that it is at an angle of 45° to the ground for 24 hours; odor stimulation: attach a test tube filled with mothballs to the cage and keep it for 12 hours; day and night reversal: avoid light from 8:00 to 20:00, and turn on lights from 20:00 to 8:00 the next day; unfamiliar object stimulation: place brushes, stones, rags, etc. in the cage and take them out after 24 hours; tail clamping: clamp the tail with a clamp 1 cm away from the base for 1.5 minutes, repeat twice.

[0211] 2.2 Behavioral evaluation

[0212] 2.2.1 Open field experiment

[0213] The rat to be tested was placed in the center of an open field box with a black bottom. The open field box was divided into nine squares with equal area, the center was the central area, and the rest were the peripheral areas. The total distance of the rat to be tested moving freely within 6 minutes was recorded and tracked using an animal behavior analysis system.

[0214] 2.2.2 Sugar water preference experiment

[0215] The sucrose water preference training and test were performed according to the reference method, and the sucrose water preference rate of each rat was calculated according to the formula: sucrose water preference rate (%) = (sucrose water consumption ÷ total liquid consumption) × 100%.

[0216] 2.2.3 Forced swimming test

[0217] The rats were gently placed in a transparent glass cylinder with a water temperature of 23±1℃ and a water depth of about 40cm. The forced swimming state of the rats was recorded with a software camera for 6 minutes, and the suspended immobility time of the rats within the next 4 minutes was analyzed.

[0218] 2.3 Thyroid ultrasound examination

[0219] The mouse neck was depilated to fully expose the examination site. After respiratory anesthesia, ultrasonic coupling agent was applied. The probe was used to perform continuous transverse scans along the long axis of the neck starting from the mandible. After the transverse scan, the probe was turned to a direction parallel to the long axis of the neck, and continuous longitudinal scans were performed from the middle of the neck to both sides. The transverse and longitudinal images of the thyroid gland were recorded respectively, and the inner and outer diameters, anterior-posterior diameters, and head-to-tail diameters of the left and right thyroid glands were measured respectively, and the volumes of the left and right thyroid glands were calculated.

[0220] 2.4 Thyroid morphology and organ index determination

[0221] The pictures in the camera were exported, grouped and classified, and the thyroid morphology was observed. The weights of the thyroid tissues on the left and right sides of the same rat were added together to obtain the total weight of the thyroid tissues, and the thyroid index was calculated using the formula. The calculation formula is as follows: thyroid index (mg / g) = thyroid weight (mg) / body weight (g).

[0222] 2.5 Thyroid tissue pathological observation

[0223] Thyroid tissue specimens were fixed in 4% paraformaldehyde for 48 h, then graded in ethanol and xylene, embedded in paraffin, cut into 3-μm-thick sections, and stained with hematoxylin and eosin. The pathological changes of thyroid tissue were observed using the HS6 automatic section analysis system.

[0224] 2.6 Serum thyroid hormone level determination

[0225] After collecting serum, the operation was strictly carried out according to the instructions of the Elisa kit to calculate the levels of FT3, FT4 and TSH in serum.

[0226] 2.7 Determination of IL-1β, IL-6 and TNF-α protein levels in thyroid tissue

[0227] Cut the thyroid tissue into small pieces and place them in a grinding tube. Add 10 times the amount of 1×PBS and grind thoroughly to prepare tissue homogenate. Centrifuge the tissue homogenate at 5000×g for 5 min at 4°C to obtain the supernatant. Take an appropriate amount of supernatant and perform the experiment strictly according to the instructions of the Elisa kit to detect and calculate the protein content of IL-1β, IL-6 and TNF-α in thyroid tissue.

[0228] 2.8 Statistical analysis

[0229] All data were statistically analyzed and plotted using Graphad Prism 8.02 software. The results were expressed as mean ± standard deviation (Mean ± SD). One-way analysis of variance was used for comparison of means between groups, and SNK-q test was used for pairwise comparison. P < 0.05 was considered statistically significant.

[0230] 3 Experimental results

[0231] 3.1 Behavioral evaluation

[0232] Compared with the blank group, the total movement distance of the rats in the model group in the open field was significantly shortened, the preference rate for sugar water was significantly reduced, and the swimming suspension time was significantly prolonged (P < 0.01), indicating that the liver qi stagnation syndrome model was successfully prepared and obvious depression appeared. Compared with the model group, there was no significant difference in the total movement distance, sugar water preference rate and swimming suspension time of the rats in the positive drug methyl lemon solution group in the open field (P > 0.05), indicating that although the positive drug methyl lemon solution has a certain improvement effect on depression, the effect is not obvious; the medium and high dose groups of the Chinese medicine gel of the present invention can significantly prolong the total movement distance of rats in the open field, increase the sugar water preference rate, and shorten the swimming suspension time (P < 0.05), indicating that the Chinese medicine gel of the present invention can significantly improve the depression of rats, and has a certain dose dependence, as shown in Fig. Figure 7 .

[0233] 3.2 Thyroid ultrasound examination

[0234] Ultrasound images showed that the thyroid gland was butterfly-shaped in cross section and triangle-shaped, narrow at the top and wide at the bottom in longitudinal section. The thyroid parenchyma of rats in the blank group had uniform echoes and clear boundaries, and the inner and outer diameters, anterior-posterior diameters, and head-to-tail diameters were all small. The glands of rats in the model group showed obvious asymmetric enlargement on both sides, with abnormal tissues, blurred and irregular boundaries, and common strong echoes of punctate or strip-like fibrous hyperplasia caused by nodular lesions. By comparing the blank group with the model group, the ultrasonic images showed that the rat model of thyroid nodules with liver qi stagnation syndrome was successfully prepared. Rats in the positive drug methyl lemon solution group and the medium and high dose groups of the Chinese medicine gel of the present invention showed relatively obvious therapeutic effects. The inner and outer diameters, anterior-posterior diameters, and head-to-tail diameters of the thyroid glands were significantly reduced, the boundaries were relatively clear and relatively regular, the thyroid gland echoes were relatively uniform, and the nodular lesions were significantly improved. Figure 8 and Fig. 9 .

[0235] Compared with the blank group, the volume of the left and right thyroid glands of the rats in the model group increased significantly (P < 0.01). Compared with the model group, there was no significant difference in the volume of the left and right thyroid glands of the rats in the low-dose group of the Chinese medicine gel of the present invention (P > 0.05); the volume of the left and right thyroid glands of the rats in the positive drug methyl lemon solution group and the medium and high-dose groups of the Chinese medicine gel of the present invention decreased significantly (P < 0.01). The therapeutic effect of the Chinese medicine gel of the present invention showed a certain dose dependence, as shown in Figure 2. Fig.10 .

[0236] 3.3 Thyroid morphology and organ index determination

[0237] The left and right lobes of the thyroid gland of the rats in the blank group were smaller in size and pink in color. The left and right lobes of the thyroid gland of the rats in the model group were significantly enlarged, and the tissues were full of blood streaks, and the colors were dark red and purple. Comparison between the blank group and the model group showed that the rat model of thyroid nodule liver qi stagnation syndrome was successfully prepared in terms of apparent morphology. The rats in the positive drug methyl lemon solution group and the high-dose group of the Chinese medicine gel of the present invention showed good therapeutic effects, and the enlargement of the left and right lobes of the thyroid gland was significantly reduced. Blood streaks were occasionally seen in the tissues, and the color was mainly light red. The medium-dose group of the Chinese medicine gel of the present invention had a certain therapeutic effect, and the enlargement of the thyroid tissue of some rats was reduced. The thyroid gland of the rats in the low-dose group of the Chinese medicine gel of the present invention was relatively severe, and blood streaks were visible in the tissues, and the color was mainly dark red. Fig.11 .

[0238] Compared with the blank group, the thyroid index of the rats in the model group increased significantly (P < 0.01). Compared with the model group, the thyroid index of the rats in the positive drug methyl lemon solution group and the high-dose group of the Chinese medicine gel of the present invention decreased significantly (P < 0.05); there was no significant difference in the thyroid index of the rats in the low- and medium-dose groups of the Chinese medicine gel of the present invention. The therapeutic effect of the Chinese medicine gel of the present invention showed a certain dose dependence, as shown in Figure 2. Fig.12 .

[0239] 3.4 Thyroid histopathological observation

[0240] There are a large number of round or oval follicles in the thyroid gland of the blank group rats, which are of medium size and regular shape. The epithelial cells are cubic or flat and arranged regularly. There is no abnormal nodular hyperplasia in the loose connective tissue between cells and follicles; the follicles in the model group are over-expanded, the number is small and the arrangement is irregular, the epithelial cells are highly nodular hyperplasia, and the boundaries between follicles are blurred. By comparing the blank group with the model group, it is shown from the histopathological aspect that the thyroid nodule liver qi stagnation rat model is successfully prepared. The follicles in the thyroid gland of the rats in the low-dose group of the Chinese medicine gel of the present invention are uneven in size and irregular in shape, and the epithelial cells are obviously hypertrophic and hyperplastic; the medium-dose group of the Chinese medicine gel of the present invention shows a certain therapeutic effect, the number of follicles is relatively increased, but the shape is still irregular, and the degree of nodular hyperplasia of follicular epithelial cells is relatively weakened. The positive drug methyl lemon solution group and the high-dose group of the Chinese medicine gel of the present invention showed good therapeutic effects. The shape and arrangement of the follicles were relatively regular, the size was relatively uniform, the loose connective tissue between cells and follicles had almost no abnormal nodular hyperplasia, the boundaries between follicles were relatively clear, the thyroid tissue morphology basically returned to normal, and the degree of inflammatory cell infiltration was significantly reduced. In addition, the therapeutic effect of the Chinese medicine gel of the present invention showed a certain dose dependence, as shown in FIG. Fig.13 .

[0241] 3.5 Serum thyroid hormone level determination

[0242] Compared with the blank group, the FT3 and FT4 levels in the serum of the rats in the model group were significantly decreased, and the TSH level was significantly increased (P < 0.01). Compared with the model group, the FT3 and FT4 levels in the serum of the rats in the low, medium and high dose groups of the Chinese medicine gel of the present invention were significantly increased (P < 0.05), and the TSH level in the serum of the rats in the medium and high dose groups of the Chinese medicine gel of the present invention was significantly decreased (P < 0.05), and the Chinese medicine gel of the present invention has a certain dose dependence in regulating the levels of FT3, FT4 and TSH; there was no significant difference in the FT3, FT4 and TSH levels in the serum of the rats in the positive drug methyl lemon solution group compared with the model group (P > 0.05). Fig.14 .

[0243] 3.6 Determination of IL-1β, IL-6 and TNF-α protein levels in thyroid tissue

[0244] Compared with the blank group, the protein contents of IL-1β, IL-6 and TNF-α in the thyroid tissue of the rats in the model group were significantly increased (P < 0.01). Compared with the model group, the protein contents of IL-1β, IL-6 and TNF-α in the thyroid tissue of the rats in the low, medium and high dose groups of the Chinese medicine gel of the present invention were significantly reduced (P < 0.05), and were dose-dependent; the protein contents of IL-1β, IL-6 and TNF-α in the thyroid tissue of the rats in the positive drug methyl lemon solution group were significantly reduced (P < 0.01). Fig.15 .

[0245] Example 6: Observation on the clinical efficacy of Chinese medicine gel for treating thyroid nodules from the perspective of liver

[0246] 1 Research subjects

[0247] Patients with thyroid nodules who met the inclusion criteria and were willing to be treated and were recruited from Jinan Central Hospital from December 2022 to December 2023 were selected.

[0248] 1.1 Inclusion criteria

[0249] It was developed with reference to the U.S. "Guidelines for the Management of Adult Patients with Thyroid Nodules and Differentiated Thyroid Cancer" (2015 edition) and the Chinese "Guidelines for the Diagnosis and Treatment of Thyroid Nodules and Differentiated Thyroid Cancer" (2012 edition).

[0250] (1) Thyroid ultrasound showed TI-RADS category 3, 4a / 4b;

[0251] (2) Patients with nodules larger than 1 cm in maximum diameter need to undergo fine needle aspiration biopsy of the thyroid nodule, and the result must be benign. Those who refuse the examination must sign a warning statement;

[0252] (3) three thyroid function tests (TSH, FT3, FT4) and three thyroid antibodies (TPOAb, TGAb, TRAb) were negative;

[0253] (4) Normal liver and kidney functions.

[0254] 1.2 Exclusion criteria

[0255] (1) Those who are taking medication for thyroid disease;

[0256] (2) Pregnant or breastfeeding women;

[0257] (3) Those who are currently participating in other clinical trials;

[0258] (4) Patients with a history of thyroid surgery;

[0259] (5) People with abnormal liver and kidney function and allergic constitution.

[0260] 1.3 Case exclusion and dropout criteria

[0261] (1) Those with severe allergies;

[0262] (2) Those who are unwilling to continue to cooperate with observation or adopt other treatments;

[0263] (3) Participating in other clinical trials during the study period;

[0264] (4) The researcher considers that the person is not suitable to continue participating in the study.

[0265] 2 Drug intervention methods

[0266] The patient cleaned the neck skin, took an appropriate amount of gel and applied it to the thyroid skin surface of the neck, and gently massaged to promote absorption, 2-3 times a day, and the intervention course was 2 months (60 days).

[0267] 3. Efficacy observation indexes

[0268] ① The maximum diameter of the thyroid nodule. The maximum diameter of the thyroid nodule is measured by thyroid ultrasound as the measurement standard, and is measured once before treatment and 2 months (60 days) after treatment;

[0269] ② TI-RADS classification of thyroid nodules. The TI-RADS classification of thyroid nodules was determined by thyroid ultrasound, once before treatment and 2 months (60 days) after treatment;

[0270] ③ Clinical symptom score: According to the patient's own symptoms, the score is evaluated once before treatment and 2 months (60 days) after treatment.

[0271] 4. Criteria for determining efficacy

[0272] 4.1 Main efficacy criteria

[0273] Changes in the maximum diameter of the nodule:

[0274] Thyroid ultrasound examinations were performed before treatment and 2 months (60 days) after medication, and the effective rate was calculated based on the change in the maximum diameter of the nodule.

[0275] ① Cured: Ultrasound shows that the maximum diameter of the nodule is reduced by ≥90% or disappears;

[0276] ② Significantly effective: Ultrasound showed that the maximum diameter of the nodule was reduced by <90%;

[0277] ③ Ineffective: Ultrasound shows that the maximum diameter of the nodule remains unchanged or even increases.

[0278] Total effective rate = (number of cured cases + number of markedly effective cases) / total number of cases × 100%

[0279] 4.2 Secondary efficacy criteria

[0280] Changes in TI-RADS classification of nodules:

[0281] Thyroid ultrasound examinations were performed before treatment and 2 months (60 days) after medication, and the effective rate was calculated based on the changes in TI-RADS classification of ultrasound results.

[0282] ① Cured: Ultrasound shows TI-RADS classification as category 2 or normal thyroid tissue;

[0283] ② Significantly effective: TI-RADS classification is reduced but remains at category 3 or 4a;

[0284] ③ Invalid: TI-RADS classification remains unchanged or increases.

[0285] Total effective rate = (number of cured cases + number of markedly effective cases) / total number of cases × 100%

[0286] Changes in clinical symptom scores:

[0287] According to the Guiding Principles for Clinical Research of New Chinese Medicines (2002 edition), the main symptoms were rated as 0, 2, 4, and 6 points, and the secondary symptoms were rated as 0, 1, 2, and 3 points. The score scale was filled in before and after treatment, and the changes in the total score were analyzed.

[0288] ① Cured: total score reduction rate ≥ 90%;

[0289] ② Significantly effective: total score reduction rate 30% to 90%;

[0290] ③ Invalid: The total score reduction rate is less than 30%.

[0291] Total score reduction rate = (total score before treatment - total score after treatment) / total score before treatment × 100%;

[0292] 5 Research Results

[0293] A total of 65 patients were included, of whom 3 withdrew voluntarily, 1 had poor compliance, 2 chose surgical treatment, and 1 was lost to follow-up, all of whom were excluded. Finally, 58 patients completed the full cycle of treatment. The maximum diameter reduction, TI-RADS classification changes, and clinical symptom score changes of the 58 patients who completed the full cycle of treatment were analyzed. The specific results are shown in Table 22. The representative results of thyroid nodule ultrasound examination in clinical practice are shown in Fig.16 , Fig.17 .

[0294] Table 22 The reduction in the maximum diameter of thyroid nodules, changes in TI-RADS classification and clinical symptom scores of patients after treatment with the Chinese medicine gel

[0295]

Claims

1. A Chinese medicine composition for treating thyroid nodules from the perspective of the liver, characterized in that: The raw material composition is as follows, in parts by weight: 15-25 parts of Prunella Vinegar, 10-20 parts of Bupleurum Root, 8-15 parts of Curcuma Atractylodes Rhizome, 8-15 parts of Forsythia suspensa, 5-15 parts of Fritillaria thunbergii, 5-15 parts of Paeonia suffruticosa, 5-8 parts of Scutellaria baicalensis, 5-10 parts of Oyster, 3-8 parts of Menthus Fructus and 3-8 parts of Borneol.

2. A Chinese medicine gel for treating thyroid nodules from the perspective of the liver, characterized in that: The extract of the Chinese medicine composition according to claim 1 is used; the raw materials of the Chinese medicine gel are composed of: 40 parts of the extract of the Chinese medicine composition, 1.5 parts of carbomer, 10 parts of glycerol, 0.5 parts of Tween-80, 2 parts of azone and 0.1 parts of ethyl hydroxybenzoate, 100 parts of purified water are added, and the pH is adjusted to 7.5 with triethanolamine.

3. The method for preparing the Chinese medicine gel according to claim 2, characterized in that: The steps of preparing the extract of the Chinese medicine composition are: (1) weighing the raw materials of the Chinese medicine composition according to proportion and setting aside; (2) adding purified water to the mint, slowly heating to boiling, and keeping it at a slight boil for 5 hours, then stopping the heating, leaving it for a while, collecting the volatile oil, and obtaining the mint volatile oil for later use; (3) soaking the peony bark in purified water, extracting it by steam distillation, and collecting the distillate; The distillate is placed at room temperature, refrigerated for crystallization, filtered, and dried under reduced pressure to obtain drug crystals; the drug crystals are mixed with borneol, ground, added with 95% ethanol to dissolve, and then peppermint volatile oil is dissolved therein to obtain solution A; (4) adding Forsythia suspensa, Prunella vulgaris, Scutellaria baicalensis, Bupleurum chinense and Oyster to purified water for decoction, concentrating, precipitating with alcohol, standing overnight and centrifuging to obtain a supernatant solution B; (5) extracting Curcuma zedoaria and Fritillaria thunbergii with ethanol, filtering, and mixing the filtrate to obtain solution C; mixing the supernatant solution B and solution C, concentrating, adding solution A, and then adding 95% ethanol to make up the volume to obtain an extract of the traditional Chinese medicine composition; (6) Weighing the raw materials of the Chinese medicine gel according to proportion; (7) After the carbomer is swollen with purified water, triethanolamine is added to adjust the pH to 7.5, and the mixture is aged to obtain a matrix I; the extract of the Chinese medicine composition, glycerol, Tween-80, azone and ethyl hydroxybenzoate are fully mixed and dissolved to obtain a composition II; the matrix I and the composition II are mixed, purified water is added to make up the weight, and the mixture is fully stirred to obtain a Chinese medicine gel.

4. The method for preparing the Chinese medicine gel according to claim 3, characterized in that: In step (2), the mass ratio of mint to purified water is 1:

8.

5. The method for preparing the Chinese medicine gel according to claim 3, characterized in that: In step (3), the mass ratio of peony bark to purified water is 1:

12.

6. The method for preparing the Chinese medicine gel according to claim 3, characterized in that: In step (4), add 8 times the amount of purified water to the medicinal material by weight, and heat and reflux to extract 3 times, each time for 1 hour.

7. The method for preparing the Chinese medicine gel according to claim 3, characterized in that: In step (5), add 6 times the weight of the medicinal material to 60% ethanol, heat and reflux to extract twice, each time for 1.5 hours.

8. The method for preparing the Chinese medicine gel according to claim 3, characterized in that: Each milliliter of the extract of the traditional Chinese medicine composition contains 1 gram of the crude drug.

9. Use of the Chinese medicine composition according to claim 1 in preparing a medicine for improving depression or treating thyroid nodules.

10. Use of the traditional Chinese medicine gel according to claim 2 in the preparation of medicines for improving depression or treating thyroid nodules.

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