CN115120691A

CN115120691A - Traditional Chinese medicine composition for improving qi deficiency and blood stasis type breast cancer perioperative immune microenvironment and life quality and application thereof

Info

Publication number: CN115120691A
Application number: CN202210701867.4A
Authority: CN
Inventors: 盛佳钰; 江科; 董梦婷; 帕提古丽·加帕尔; 袁敏佳; 李晓菲
Original assignee: Yueyang Hospital of Integrated Traditional Chinese and Western Medicine Shanghai University of TCM
Current assignee: Yueyang Hospital of Integrated Traditional Chinese and Western Medicine Shanghai University of TCM
Priority date: 2022-06-21
Filing date: 2022-06-21
Publication date: 2022-09-30
Anticipated expiration: 2042-06-21
Also published as: CN115120691B

Abstract

The invention relates to the field of traditional Chinese medicines, in particular to a traditional Chinese medicine composition for improving qi deficiency and blood stasis type breast cancer perioperative immune microenvironment and life quality and application thereof, wherein the traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 15-45 parts of astragalus root, 15-45 parts of zedoary, 5-25 parts of Chinese sage herb, 5-25 parts of codonopsis pilosula and 3-18 parts of medlar. The traditional Chinese medicine composition is used for orthotopically centering the pathogenesis of qi deficiency and blood stasis after breast cancer operation, and has the effects of tonifying qi, strengthening body resistance, promoting blood circulation and removing blood stasis; cytological experiments prove that the drug-containing serum of the 'stilbene operation prescription' can inhibit macrophage polarization to M2 type, further inhibit breast cancer MDA-MB-231 cell proliferation, and has better effect than the drug-containing serum of 'breast cancer operation rear'. Clinical experiments prove that the astragalus formula can regulate the immune microenvironment of a breast cancer patient in the perioperative period of qi deficiency and blood stasis type by reducing the level of peripheral blood M2 macrophages, and can improve the survival quality of the breast cancer patient in the perioperative period and relieve the symptoms of traditional Chinese medicine.

Description

Traditional Chinese medicine composition for improving qi deficiency and blood stasis type breast cancer perioperative immune microenvironment and life quality and application thereof

Technical Field

The invention relates to the technical field of traditional Chinese medicines, in particular to a traditional Chinese medicine composition for improving qi deficiency and blood stasis type breast cancer perioperative immune microenvironment and life quality and application thereof.

Background

Breast cancer is the most common female malignancy, accounting for about 30% of new cases of female cancer worldwide. Despite the maturity of breast cancer treatment modalities, nearly 26.5% of patients die due to treatment failure. Currently, surgical treatment is still an essential part for breast cancer patients who are clinically staged to be operable. The reasons for recurrence or metastasis of a tumor after surgery are complex, and are not only influenced by the surgical operation and the biological properties (invasion and metastasis) of the tumor itself, but also closely related to the immune microenvironment in which the tumor is located during the perioperative period (Looney M, Doran P, pocket DJ. Effect of infectious technology on mouse end growth factor C and transformation growth factor beta in mouse end grafting and metastasis for cancer [ J ] Anestheology, 2010,113(5):1118 1125.). In recent years, researches show that most of the clinical narcotic drugs including inhalation narcotic sevoflurane, isoflurane, local anesthetic lidocaine, ropivacaine, morphine and the like can inhibit the immunity of patients, so that tumor cells can avoid immune monitoring and escape, thereby increasing the tumor micrometastasis rate in the perioperative period and further improving the risk of postoperative tumor recurrence. Therefore, a treatment scheme for improving tumor immune microenvironment in the perioperative period is urgently needed to be found, so that the occurrence of immune escape can be effectively inhibited, and the prognosis can be improved. The application of the traditional Chinese medicine is the characteristics and advantages of the prevention and treatment research of breast cancer in China. A large number of literature reports and preliminary studies of subject groups all prove that the traditional Chinese medicine intervention can effectively reduce the recurrence and metastasis rate of breast cancer and prolong disease-free survival, and is closely related to the immunoregulation effect of the breast cancer. At present, the safety of the traditional Chinese medicine in perioperative period is approved. (Shengjiayu, Zhang Xinyue, Liqiong, Xue Xiaohong, discussing the influence of the postoperative course of breast cancer on the lung metastasis of the breast cancer mouse based on the change of TLR 4/NF-kB pathway in the tumor microenvironment [ J ]. Chinese traditional medicine journal, 2016,34(12):2864 plus 2867. Shengjiayu, Liu Yu, Yan Meixia, Jianke, and the meaning of the traditional Chinese medicine for the new adjuvant therapy of breast cancer [ J ]. Yunnan academy of academic, 2018,41(3):47-50+54. Yanghui, Sunxin, Liuxi, liver-soothing and kidney-tonifying formula plus and minus the influence of the immune function and the life quality of the breast cancer patient [ J ]. modern traditional Chinese and western medicine combined journal, 2020, 29; 3683 plus 3687. huge Zhi just, spring silk, Jujuanjuan, Weijin Wei Peng, Chen Peng, Zhang Shi Xianfeng Feng and Qi-benefiting decoction for the level of the relevant factors of breast cancer and the research [ J ], 2020,38(12):138-141.).

Most scholars believe that breast cancer (mammary cancer) is mainly caused by qi stagnation and blood stasis, pathogenic toxin accumulation in the interior and milk collaterals stagnation due to dysfunction of viscera and disorder of qi and blood of Chong and ren meridians caused by deficiency of vital qi, invasion of exogenous pathogenic factors, internal injury of seven emotions and the like. The deficiency of healthy qi is the key to the onset of disease, and invasion of pathogenic factors is an important condition for the disease. The national famous Chinese medicine surgery school-professor of fourth generation of Gu's science heritage LudeMing proposes the view of self-eliminating of healthy qi and pathogenic factors and assisting tumor elimination in treating breast cancer, establishes the treatment principles of strengthening healthy qi to eliminate pathogenic factors and regulating Chong and conception vessels, establishes the postoperative treatment method of breast cancer of tonifying qi and nourishing yin, regulating Chong and conception vessels and detoxifying, and establishes the postoperative prescription of breast cancer. The formula follows the basic treatment rule of strengthening body resistance and eliminating pathogenic factors, and adopts astragalus root, radix codonopsitis, epimedium, dogwood fruit, tuckahoe, largehead atractylodes rhizome and medlar to tonify qi and spleen, warm kidney and strengthen yang, tonify both the first and the next day, and can also regulate thoroughfare and conception vessels to strengthen body resistance and consolidate constitution; furthermore, it is combined with Shi Ying, E Zhu and E He to activate blood and resolve stasis, remove toxicity and prevent cancer, so it can prevent the residual toxicity from escaping. The prescription is widely applied to the postoperative convalescence of breast cancer, namely 0-5 years after the operation and the prescription chemotherapy treatment are finished. The previous research finds that the formula can improve the physiological, emotional and cognitive functions of patients and improve the life quality; but also can delay the time of recurrence and metastasis of breast cancer patients, improve the disease-free survival rate and reduce the disease death rate. (Wanhua, Wu Xuehuanqing, Fried Hui, Dong Jiarong, Liusheng, Ludeming. "Breast cancer after surgery" clinical research on the quality of life of Breast cancer patients [ J ]. Shanghai journal of medicine, 2004(9):28-31.Nan Pen, Mingwei Yu, Guiwang Yang, Qi Fu, Yongmei Xu, Jie Yu, Qiang Liu, Chen Li, Weiru Xu, Yi Zhang, Cong Maase, Lin Yang, Rencun Yu, Xiaomin Wang.Effects of the Chinese Yin Shen Jian Gununles on granules, restriction-tissue, and said tissue, Xiwanwei Juliang, Huang Julian, Julian, Xiwang Julian, Julian, Juliang, Julian, Tian, Yiwanhua, Yiwang Yang, Yilian, Yiganshan, Yigane, Hainan Julian, Wan Julian, Wan, Julian, Wan, Julian, Wan Julian, Wan, Julian, Wan, Julian, Wan, Julian, Wan, U, Wan, U, Wan, U, Wan, U, Wan, U, Wan, U, impact of non-diseased and Total survival after 10 years post-operative Breast cancer in von Jiamei, Gaoyanqian, Shaoshu, Sun bright, Breast cancer post-operative team [ J ] J.J.Utility Med., 2021, 37(15):1988-1992.)

However, no report is found on the traditional Chinese medicine composition aiming at the qi deficiency and blood stasis type breast cancer in the perioperative period at present.

Disclosure of Invention

The invention aims to provide a traditional Chinese medicine composition for improving qi deficiency and blood stasis type breast cancer perioperative immune microenvironment and life quality and application thereof, aiming at the defects in the prior art.

The blood of tumor patients is often in a high coagulation state (MiXK, Liu QR, Zhu L, SangMX, Guo LR, Shann BE.mechanism of the high coagulation state of Breast cancer tissue factor [ J ]. Eur Rev Med Pharmacol Sci,2017,21(9):2167-2171.), and the stress reaction caused by the operation also can promote the blood coagulation function of the patients to be further activated, thereby causing the occurrence of postoperative venous thrombosis and pulmonary embolism of lower limbs. In addition, the traditional Chinese medicine theory considers that the operation trauma consumes qi and blood, qi deficiency can not lead to blood circulation, blood circulation is not smooth, and venation is stagnant, so the qi deficiency and blood stasis syndrome is the main syndrome type of the breast cancer patient in the perioperative period, and the symptoms of qi deficiency and blood stasis are more severe than those of the breast cancer patient in the postoperative recovery period, so that the difference between the medical syndrome type of the breast cancer patient in the perioperative period and the medical syndrome type of the breast cancer patient in the postoperative recovery period is larger, and the breast cancer postoperative prescription is not suitable for the qi deficiency and blood stasis syndrome of the breast cancer patient in the perioperative period. The traditional Chinese medicine composition provided by the invention is derived from the postoperative of breast cancer, mainly aims at the syndrome of qi deficiency and blood stasis of patients in the perioperative period of breast cancer, has a formula principle and a treatment method different from the postoperative formula of breast cancer, and purposely adjusts the dosage of the medicine, so that a 'astragalus formula' is formed with good effect, and the immune microenvironment and the quality of life in the perioperative period of breast cancer are improved for the patients with qi deficiency and blood stasis syndrome in the perioperative period of breast cancer.

The invention provides a traditional Chinese medicine composition for improving immune microenvironment and quality of life in a breast cancer perioperative period, which is prepared from the following raw material medicines in parts by weight: 15-45 parts of astragalus root, 15-45 parts of zedoary, 5-25 parts of Chinese sage herb, 5-25 parts of codonopsis pilosula and 3-18 parts of Chinese wolfberry.

Further, the perioperative period is the period of time (typically within two weeks of surgery) that the patient decides to undergo surgery, perform surgery and recover from surgery (Bollen Pinto B, Chew M, Lurai Buse G, Walder B. the concept of the positive medical to the positive medical additives A. the positive review. Eur J Anaesthesiol.2019 Dec; 36(12): 889) 903.).

Further, the traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 20-40 parts of astragalus membranaceus, 20-40 parts of curcuma zedoary, 10-20 parts of Chinese sage herb, 10-20 parts of codonopsis pilosula and 6-12 parts of medlar.

In a preferred embodiment of the invention, the traditional Chinese medicine composition is prepared from the following raw medicines in parts by weight: 30 parts of astragalus, 30 parts of curcuma zedoary, 15 parts of Chinese sage herb, 15 parts of codonopsis pilosula and 9 parts of medlar.

The invention provides an application of the traditional Chinese medicine composition in preparation of a medicine for improving qi deficiency and blood stasis type breast cancer perioperative immune microenvironment and life quality.

Further, the improvement of the qi deficiency and blood stasis type breast cancer perioperative immune microenvironment refers to inhibition of macrophage M2 type polarization and inhibition of breast cancer immune escape.

Furthermore, the improvement of the life quality refers to the regulation of emotional management disorder caused by emotional disorder after the operation of the breast cancer patient, the improvement of qi deficiency and blood stasis symptoms such as qi deficiency fatigue, insomnia, spontaneous perspiration and the like, and the improvement of the life quality of the patient.

The third aspect of the invention provides a pharmaceutical preparation using the above traditional Chinese medicine composition for improving qi deficiency and blood stasis type breast cancer perioperative immune microenvironment and quality of life as an active ingredient, wherein the pharmaceutical preparation is prepared into a pharmaceutically common dosage form by adopting a conventional preparation method in the field.

Further, the medicinal preparation is decoction, pills, tablets, mixture, capsules, granules, powder and paste.

Furthermore, the pharmaceutical preparation also comprises pharmaceutically acceptable auxiliary materials. Furthermore, the pharmaceutically acceptable auxiliary materials comprise emulsifying agents, filling agents, binding agents, wetting agents, disintegrating agents, absorption promoters, flavoring agents, coloring agents and cosolvent.

The invention has the advantages that:

the traditional Chinese medicine composition is a clinical proved formula, wherein astragalus membranaceus in the formula has the effects of tonifying qi and strengthening body resistance, and curcuma zedoary has the effects of removing blood stasis and activating blood circulation, and is a monarch drug; the ministerial drug, namely the salvia chinensis, has the effects of activating blood and dredging collaterals, and the codonopsis pilosula has the effects of tonifying middle-jiao, Qi and spleen and gaining monarch drugs; the medlar is used for promoting the production of body fluid and nourishing yin, and the medicines have the effects of tonifying qi and strengthening body resistance, and activating blood and dissolving stasis. Cytological experiments prove that the drug-containing serum of the 'stilbene operation prescription' can inhibit macrophage polarization to M2 type, further inhibit breast cancer MDA-MB-231 cell proliferation, and has better effect than the drug-containing serum of 'breast cancer operation rear'. Clinical experiments prove that the astragalus prescription can regulate the perioperative immune microenvironment of a patient with qi deficiency and blood stasis type breast cancer by reducing the level of peripheral blood M2 macrophages, has a prominent regulating and controlling effect on peripheral blood M2 type macrophages of a patient with Luminal B type breast cancer, and can possibly become an important breakthrough of the bottleneck of Luminal B type breast cancer treatment. Meanwhile, the traditional Chinese medicine composition can also regulate emotion management disorder caused by emotional disorder after the operation of a breast cancer patient, improve symptoms of qi deficiency and blood stasis such as qi deficiency fatigue, insomnia, spontaneous perspiration and the like, improve the life quality of the patient, and relieve traditional Chinese medicine symptoms.

Drawings

FIG. 1 is a schematic view of the study procedure of the clinical trial of example 19.

FIG. 2 shows the ratio of macrophage M1/M2 after the drug-containing serum is used for treating simple macrophages and MDA-MB-231 cell and macrophage co-culture system respectively; note: comparison with corresponding macrophage + drug serogroup ^* P<0.05， ^** P<0.05; comparison between two groups ^# P<0.05。

FIG. 3 comparison between the indexes before and after intervention in the Qishu group and the placebo group; a: CD14 before intervention of Qishu prescription ⁺ And CD14 ^- Macrophages are at CD3 ^- CD19 ^- PM-2K ⁺ Percentage of CD14 cells; b: CD14 after intervention of Qishu prescription ⁺ And CD14 ^- Macrophages are at CD3 ^- CD19 ^- PM-2K ⁺ Percentage of CD14 cells; c: pre-intervention CD14 for placebo group ⁺ And CD14 ^- Macrophage cell proliferation in CD3 ^- CD19 ^- PM-2K ⁺ Percentage of CD14 cells; d: CD14 after placebo group intervention ⁺ And CD14 ^- Macrophage cell proliferation in CD3 ^- CD19 ^- PM-2K ⁺ Percentage of CD14 cells; e: IL-10 and CD14 between the groups before and after intervention of the Qishu group and the placebo group ⁺ Jointly detecting a comparison condition; f: IL-10 and CD14 in the group before and after intervention in the Qishu prescription group and placebo group ⁺ Jointly detecting a comparison condition; note: ns: no significant statistical significance; *: p is less than 0.05; **: p is less than 0.01; ***: p is less than 0.001.

Detailed Description

The following examples are provided to illustrate specific embodiments of the present invention.

Example 1: the Chinese medicinal composition of the invention

The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 40 parts of astragalus root, 40 parts of zedoary, 20 parts of Chinese sage herb, 20 parts of codonopsis pilosula and 12 parts of medlar.

Example 2: the invention relates to a Chinese medicinal composition

The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 30 parts of astragalus, 30 parts of curcuma zedoary, 15 parts of Chinese sage herb, 15 parts of codonopsis pilosula and 9 parts of medlar.

Example 3: the invention relates to a Chinese medicinal composition

The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 20 parts of astragalus membranaceus, 20 parts of curcuma zedoary, 10 parts of Chinese sage herb, 10 parts of codonopsis pilosula and 6 parts of medlar.

Example 4: the Chinese medicine composition of the invention

The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 30 parts of astragalus, 30 parts of curcuma zedoary, 20 parts of Chinese sage herb, 20 parts of codonopsis pilosula and 12 parts of medlar.

Example 5: the Chinese medicine composition of the invention

The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 30 parts of astragalus, 30 parts of curcuma zedoary, 10 parts of Chinese sage herb, 10 parts of codonopsis pilosula and 6 parts of medlar.

Example 6: the Chinese medicine composition of the invention

The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 40 parts of astragalus, 40 parts of curcuma zedoary, 15 parts of Chinese sage herb, 15 parts of codonopsis pilosula and 9 parts of medlar.

Example 7: the Chinese medicine composition of the invention

The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 20 parts of astragalus, 20 parts of curcuma zedoary, 15 parts of Chinese sage herb, 15 parts of codonopsis pilosula and 9 parts of medlar.

Example 8: the Chinese medicine composition of the invention (eight)

The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 30 parts of astragalus, 20 parts of curcuma zedoary, 15 parts of Chinese sage herb, 15 parts of codonopsis pilosula and 9 parts of medlar.

Example 9: the Chinese medicine composition of the invention (nine)

The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 20 parts of astragalus, 30 parts of curcuma zedoary, 15 parts of Chinese sage herb, 15 parts of codonopsis pilosula and 9 parts of medlar.

Example 10: the Chinese medicinal composition of the invention (ten)

The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 30 parts of astragalus, 30 parts of curcuma zedoary, 20 parts of Chinese sage herb, 10 parts of codonopsis pilosula and 9 parts of medlar.

Example 11: the Chinese medicinal composition of the invention (eleven)

The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 30 parts of astragalus, 30 parts of curcuma zedoary, 10 parts of Chinese sage herb, 20 parts of codonopsis pilosula and 9 parts of medlar.

Example 12: the Chinese medicinal composition of the invention (twelve)

The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 15 parts of astragalus, 15 parts of curcuma zedoary, 5 parts of Chinese sage herb, 5 parts of codonopsis pilosula and 3 parts of medlar.

Example 13: the Chinese medicinal composition of the invention (thirteen)

The traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 45 parts of astragalus root, 45 parts of zedoary, 25 parts of Chinese sage herb, 25 parts of codonopsis pilosula and 18 parts of medlar.

Example 14: preparation of decoction of Chinese medicinal composition of the invention

Weighing the raw materials according to the weight part ratio of any one of the embodiments 1 to 13, crushing, adding a proper amount of water, and decocting by a conventional method.

Example 15: preparation of the Chinese medicinal composition mixture

Weighing the raw materials according to the weight part ratio of any one of embodiments 1-13, mixing, crushing, adding 8 times and 10 times of water, decocting twice for 1 hour each time, filtering respectively, combining the filtrates, concentrating to a relative density of 1.20(80-85 ℃), cooling, adding 3 times of ethanol, stirring, standing, filtering the supernatant, steaming the filtrate without alcohol smell, standing, and concentrating the supernatant; adding appropriate pharmaceutical adjuvants (white sugar, Mel, benzyl propionic acid or hydroxy phenyl ethanol, etc.), and making into mixture.

Example 16: preparation of the pharmaceutical composition pill of the invention

Weighing the raw materials according to the weight part ratio of any one of embodiments 1-13, crushing into fine powder, and sieving with a 80-mesh sieve for later use; weighing a certain amount of honey, heating in an evaporation dish until the honey is boiled (if magazines can be filtered), continuously refining to obtain refined honey, and removing floating foams; mixing refined honey and medicinal powder according to the proportion of 1: 1, fully and uniformly mixing; placing the well-mixed dough-like soft material for a certain time; the pills are manually made into smooth and spherical pills and are wrapped by wax paper.

Example 17: preparation of the powder of the pharmaceutical composition of the present invention

Weighing the raw materials according to the weight part ratio of any one of embodiments 1-13, mixing, crushing into fine powder, sieving with a 80-mesh sieve, and packaging with wax paper.

Example 18: preparation of the pharmaceutical composition paste of the present invention

Weighing the raw materials according to the weight part ratio of any one of embodiments 1-13, mixing, crushing, adding 8 times and 10 times of water, decocting twice for 1 hour each time, filtering respectively, combining the filtrates, concentrating to a relative density of 1.20(80-85 ℃), cooling, adding 3 times of ethanol, stirring, standing, filtering the supernatant, steaming the filtrate without alcohol smell, standing, concentrating the supernatant under reduced pressure to obtain an extract, dripping the extract on mulberry paper without infiltration water, and sealing in a sterile bottle.

Example 19: in vitro cell assay of the Chinese medicinal composition of the present invention

1 materials

1.1 animals and cell lines

Male SD rats, SPF grade, 6 weeks old, body mass (200. + -. 10) g, purchased from Shanghai Stark laboratory animals Ltd, animal license number SCXK (Shanghai) 2013-. The food is raised in an SPF-level animal room of an animal experiment center of Yueyang Hospital affiliated to Shanghai medical university, the temperature is 23 +/-3 ℃, the relative humidity is 35-45%, the illumination is 12 hours, and drinking water can be freely taken.

MDA-MB-231 human breast cancer cells, THP-1 human monocytes were purchased from Shanghai Life sciences institute of Chinese academy of sciences. MDA-MB-231 cells were cultured in L-15 medium containing 10% Fetal Bovine Serum (FBS); THP-1 cells were cultured in RPMI 1640 medium containing 10% FBS.

1.2 reagents and instruments

Phorbol ester (PMA), thiazole blue (MTT), pancreatin cell digest (Sigma, usa); CD86-APC human (Miltenyi, Germany, Cat. 305206); APC anti-human CD206 (Biolegend, USA, with a code of 321110). An ELX800 full-automatic enzyme labeling instrument (Bio-Tek, USA), a C6 analysis type flow cytometer (BD Biosciences, USA), and a low-temperature centrifuge (Eppendorf).

1.3 medicaments and preparation

Decoction pieces prepared in a 'astragalus technology prescription' (30 g of astragalus mongholicus, 30g of curcuma zedoary, 15g of salvia chinensis, 15g of codonopsis pilosula and 9g of Chinese wolfberry) and 'breast cancer postoperative' (15 g of astragalus mongholicus, 10g of codonopsis pilosula, 10g of bighead atractylodes rhizome, 12g of adenophora tetraphylla, 12g of wolfberry fruit, 15g of epimedium herb, 9g of dogwood, 12g of cistanche deserticola, 15g of salvia chinensis, 9g of honeycomb, 9g of curcuma zedoary and 12g of poria cocos) are purchased from a Yueyang Chinese-western medicine combination hospital affiliated to Shanghai-Zhongyao medicine university. Decocting by conventional method, soaking in 10 times of water for 30min, boiling with strong fire, decocting with slow fire for 40min, filtering the decoction with gauze, adding 8 times of water, boiling with strong fire, decocting with slow fire for 40min, and filtering the decoction with gauze. Mixing the liquid medicines for 2 times, concentrating the liquid medicine of the 'Qishufang' to 1.98g/mL of the raw medicinal material in water bath, concentrating the liquid medicine of the 'Ruma shufang' to 1.40g/mL of the raw medicinal material, subpackaging by using a 50mL centrifuge tube, and storing in a refrigerator at 4 ℃.

1.4 preparation of drug serum

Experimental rats were randomly divided into 2 groups of 5 rats each. The rats in the group of the 'Qishu formula' are subjected to intragastric administration by 15mL/kg of the decoction of the 'Qishu formula', and the blank control group is subjected to intragastric administration by equal volume of distilled water for 2 times/day and 3 days continuously. 1h after the last administration, performing ether anesthesia, taking blood from abdominal aorta, collecting by using a sterile centrifuge tube, standing for 2h at 4 ℃, centrifuging for 15min at 3000r/min at 4 ℃, mixing and combining the serum of the rat drug of the same group, inactivating for 30min in a 56 ℃ constant temperature water bath, filtering and sterilizing by using a 0.22 mu m microporous filter membrane, subpackaging by using a freezing tube, and storing in a refrigerator at-80 ℃ for later use. Preparing into 'Qishufang' medicinal serum culture solution with final concentration of 2.5%, 5% and 10%, respectively. [ final concentration (%). serum volume/total volume x 100% ]. The "post-operative breast cancer" drug serum culture solutions were prepared in the same manner at final concentrations of 2.5%, 5%, and 10%, respectively.

2 method

2.1 detection of cell Activity by MTT colorimetric method

Collecting THP-1 human mononuclear cells in logarithmic growth phase, and adjusting cell density to 5 × 10 ⁴ one/mL of the cells were inoculated into a 96-well plate at 100. mu.L/well, PMA was added to give a final mass concentration of 50. mu.g/L, and 5% CO was added at 37 ℃ ₂ Culturing and inducing for 24h adherence in a constant temperature incubator to obtain the induced macrophage. After the macrophage adheres to the wall, the culture is continued by changing to a 'Qishu formula' containing 2.5 percent, 5 percent and 10 percent of medicine serum culture solution containing 'mammary cancer postoperative' medicine serum. Culturing for 24h and 48h, respectively adding 20 mu L of MTT (methyl thiazolyl tetrazolium) with the concentration of 5g/L into each well cell, acting for 4h at 37 ℃, discarding supernatant, respectively adding 150 mu L of dimethyl sulfoxide (DMSO) into each well, shaking for dissolving, and reading the absorbance A of each well cell at the wavelength of 570nm on an enzyme labeling instrument to detect the influence of drug-containing serum with different concentrations on the activity of macrophages, namely the stilbene operation method and the breast cancer operation method.

2.2 detection of cell proliferation by MTT colorimetric method

Macrophage induction as in item 2.1. Changing new culture solution, adding 2.5% of "Qishufang" and 2.5% of "Breast cancer postoperative" medicinal serum culture solution, and reacting for 24 hr. Sucking the medicinal supernatant, adding 2 × 10 ⁵ MDA-MB-231 cells of one/mL, 100. mu.L/well, 2 cells in a ratio of 1: 1, were co-cultured for 24 h. Inverted microscopeAnd observing the state of cells in the hole, and taking a picture. Adding 20 mu L of MTT (methyl thiazolyl tetrazolium) with the concentration of 5g/L into each hole cell, acting at 37 ℃ for 4 hours, discarding supernatant, adding 150 mu L of DMSO into each hole cell, oscillating to dissolve, reading A of each hole cell at the wavelength of 570nm on an enzyme labeling instrument, and calculating the tumor cell inhibition rate.

2.3 flow assay for macrophage polarization

Conventionally culturing THP-1 cells, collecting logarithmically grown cells, adjusting cell density to 2X 10 ⁷ After cells per mL are paved on a 6-well plate and 2mL per well and PMA induces cell adherence, 2.5 percent of medicine serum culture solution of 'Qishufang' is added. Placing at 37 ℃ with 5% CO ₂ The incubator is used for 24 h. Cells were harvested and flow stained for cell surface antigen CD86 and intracellular antigen CD206, respectively. Wherein CD 8610. mu.L of extracellular antigen CD86 antibody is added to each tube for staining, and incubation is carried out for 30min in the absence of light. Centrifuge at 300 Xg for 3min, wash 2 times, discard the supernatant, add PBS 500. mu.L to resuspend. Cell purification per tube for CD206 intracellular antibody staining&Fixative A in Permeabilization Kit, 100. mu.L/tube, protected from light at room temperature for 15 min. Centrifuge at 300 Xg for 3min, wash 1 time, discard the supernatant, add PBS 100. mu.L to resuspend. Adding the membrane-rupturing liquid B, 100 mu L/tube, CD206, 5 mu L/tube. And blowing, beating and mixing uniformly, and keeping away from light for 15min at room temperature. Centrifuging at 300 Xg for 3min, washing for 2 times, discarding the supernatant, adding 500 μ L PBS for resuspension, mixing well, protecting from light, placing on ice, and detecting with flow cytometry. The results were analyzed using FlowJo software.

3 results

3.1 Effect of drug-containing serum on macrophage growth in Astragalus prescription and Breast cancer postoperative

Compared with the blank group, 2.5 percent of the 'Qishufang' medicament serum culture solution has 24 hours and 48 hours of action, and the growth of macrophages has no obvious change; 2.5 percent of the serum culture solution of the medicine for treating the breast cancer postoperative has no obvious change in the growth of macrophages after 24 hours of action, and the growth of the macrophages is inhibited after 48 hours of action (P < 0.05). Therefore, 2.5% of the drug-containing serum selected in the experiment from the 'stilbene method' and 'breast cancer method' acts on macrophages. See table 1.

TABLE 1 Effect of drug-containing serum on macrophage proliferation after Astragalus therapy and Breast cancer therapy: (

n＝3)

Note: comparison with blank group ^* P<0.05， ^** P<0.01。

3.2 Effect of macrophages after action of drug-containing serum on MDA-MB-231 Breast cancer cell proliferation

Compared with a model group, macrophages can obviously inhibit the proliferation of MDA-MB-231 cells (P is less than 0.01) after 2.5 percent of 'stilbene operation formula' and 2.5 percent of 'breast cancer operation post' medicament-containing serum act, and the medicament-containing serum inhibiting effect of the 'stilbene operation formula' is obviously superior to that of 'breast cancer operation post'. See table 2.

TABLE 2 Effect of macrophages with drug and serum on MDA-MB-231 cell proliferation

n＝5)

Note: comparison with model group ^* P<0.01。

3.3 Effect of drug-containing serum on macrophage polarization in Astragalus prescription and Breast cancer post-operation

Drug-containing serum treatment of in vitro induced macrophages with 2.5% of "stilbene method" and 2.5% of "post-breast cancer" was found to be: the macrophage + stilbene and macrophage + breast cancer post-operative cohorts M1/M2 ratios were not significantly changed compared to the macrophage cohort (P > 0.05). And after 2.5% "stilbene operation formula" and 2.5% "breast cancer operation post" medicated serum treatment of the MDA-MB-231 cell and macrophage co-culture system, the findings are that: the macrophage + MDA-MB-231+ breast cancer postoperative formula group and the macrophage + MDA-MB-231+ stilbene operation formula group M2 macrophage ratio are lower than the macrophage + breast cancer postoperative formula group (P <0.05) and the macrophage + stilbene operation formula group (P < 0.01). The M2 macrophage ratio was significantly lower than the macrophage + stilbene surgery group (P <0.01), and the M2 macrophage ratio for the macrophage + MDA-MB-231+ stilbene surgery group was also significantly lower than the macrophage + MDA-MB-231+ breast cancer post-surgery group (P <0.05) (fig. 2).

4 discussion of

Tumor Microenvironment (TME) is important soil for the occurrence and development of breast cancer. Among TME-related cell populations, tumor-associated macrophages (TAMs) are increasingly well known to scholars for their polarizing and remodelable characteristics, and are primarily characterized by activation or remodeling into M1-type and M2-type macrophages in different tumor microenvironments. In contrast to the pro-inflammatory tumor suppressive effects of macrophages of type M1, macrophages of type M2 secrete anti-inflammatory factors such as IL-10, transforming growth factor beta, and assist tumor cells in immune escape, while releasing multiple substrates to facilitate angiogenesis and remodeling, and are important factors in adverse prognosis such as lymphoid metastasis (Steenbrunge J, Breyne K, demeyer K, De Wever O, Sanders NN, Van Den Broeck W, Colpate C, Vermeulen P, Van Laere S, Meyer E.anti-inflammatory signaling and negative breast tumor cells, media promotion specific therapy mazoparation, polarization in an innovative tissue model for triple-negative tissue center [ J ]. J Exp, Cancer cell, research 2018, Sankyr J, Sanker J, Sanchen K, German cell K, German tumor TM J, Sanders N K, Sanders N De N K, C, Sanders N K, N K, N K, N III, N, PD-1expression by structural activities and structural importance [ J ] Nature,2017,545(7655) 495-499.Lala PK, Nandi P, Majuder M.Roles of protagans in structural-associated physiology-genetics with specific reference to crack sampler [ J ] Cancer metals Rev,2018, 36937 (2-3): 384). It has been found that macrophages in the microenvironment of breast Cancer tumors are mainly composed of M2 type macrophages (Bao XW, Shi R, Zhao TY, Wang YF, Anastasov N, Rosemann M, Fang WJ. integrated analysis of single-cell RNA-seq and bulk RNA-seq un tissue promoter plus M2-like tissue promoter-associated macro expression and aggregation in TNBC [ J ] Cancer immune tissue, 2021,70(1):189 and 202.), and the present invention identifies the type of breast Cancer immune cell infiltration using the breast Cancer single cell data set GSE75688 in the GEO database, and the results are consistent with the literature reports. In order to further prove the relation between macrophage M2 polarization and breast cancer prognosis, the invention carries out CD68, CD16 and CD163 detection (respectively marking total macrophages, M1 type macrophages and M2 type macrophages) on the tissue chip of 169 postoperative patients of breast cancer by an immunohistochemical method, median follow-up is 65 months, so as to confirm the relevance between the total infiltration number of three cell groups and the patient prognosis, and the result shows that: the prognosis of the patients with high expression of CD163 is poor (P ═ 0.023), the prognosis of high expression of CD16 is good (P ═ 0.046), and CD68 has no obvious correlation with the prognosis of breast cancer (P ═ 0.283). The M2 type macrophage plays a role in the immune microenvironment of the breast cancer, and the M2 type macrophage polarization is closely related to the poor prognosis of the breast cancer patient.

The perioperative treatment of malignant tumor patients plays a significant role, and the key to the influence on prognosis is how to change the tumor immune microenvironment of breast cancer patients in perioperative period and inhibit the occurrence of immune escape. The qi-tonifying and blood-activating method represented by the 'Qishu prescription' is a main treatment rule and treatment method for breast cancer comprehensive treatment in the perioperative period in our department, and has definite clinical curative effect but unknown specific mechanism. Cytological studies found that: the in vitro cell administration of the 'Qishu prescription' can obviously reduce the proportion of M2 type macrophages in a co-culture system of the macrophages and breast cancer cells, and the effect is better than that of the breast cancer postoperative prescription. And (4) prompting: the "stilbene method" may inhibit breast cancer immune escape by altering macrophage M2 polarization.

Example 20: clinical trials of the pharmaceutical composition of the present invention

1 data and method

1.1 study object

During the period from 7 months in 2021 to 12 months in 2021, 80 cases of patients with primary breast cancer who were treated by the Yueyang Chinese and western medicine combination hospital affiliated to Shanghai medical university were selected. All the selected cases were confirmed to be breast cancer by histology or cytology. Staging criteria meet the criteria set forth in the United states Joint Committee for Cancer (American Joint Committee on Cancer, AJCC) 7 th edition regarding staging of tumors. 80 patients were all female, median age 58 years in the stilbene surgery patients, pathological type: invasive ductal carcinoma 37 cases (92.5%); invasive lobular carcinoma 3 cases (7.5%), median age 61 years in placebo patients, pathological type: invasive ductal carcinoma 38 cases (95.0%); invasive lobular carcinoma 2 cases (5.0%). All patients with breast cancer and healthy people who entered the group filled in the informed consent, and the BC patients had different clinical pathological characteristics, such as age, TNM stage, tumor size, lymph node metastasis, Estrogen Receptor (ER) status, human epidermal growth factor 2 (Her-2) status, etc., as detailed in table 3.

TABLE 3 clinical and pathological parameters of breast cancer group and healthy control group

1.2 nano-rank standard

1.2.1 inclusion criteria: the age of the female is more than or equal to 18 years and less than or equal to 80 years; secondly, the result of the histopathology is confirmed to be invasive breast malignant tumor; and the diagnosis standard of the deficiency of vital energy and blood stasis syndrome in the traditional Chinese medicine is met. Fourthly, 1 day after the operation treatment is finished; fifth, any tumor-related treatment except surgery, including radiotherapy, chemotherapy, endocrine treatment, etc., is not received; sixthly, the patient gives consent.

1.2.2 exclusion criteria: rejecting drawing blood or unable collaborators due to suffering from mental diseases and the like; combining serious diseases of a circulatory system, a respiratory system, a digestive system, a blood system, a urinary system, a nervous system and an endocrine system, and considering that serious diseases of other systems of the patient influence the research result; ③ distant metastasis of breast cancer; (iv) those known to have any other malignancy; the other clinical researchers are participated in the same period; sixthly, the patient is in pregnancy or lactation; seventhly, the traditional Chinese medicine cannot be taken due to allergy and the like; the patient regularly takes the traditional Chinese medicine for nearly three months before the experiment (taking the traditional Chinese medicine more than or equal to 5 days per week with the interruption of less than or equal to 7 days).

1.3 detection method

1.3.1 flow cytometry of peripheral blood macrophages: 5ml of anticoagulated whole blood was added to 15ml of Ficoll (tertiary amine salt of Tianjin), centrifuged at 800g for 20min, and the buffy coat was removed and washed once to isolate PBMC. The samples were blocked with 10% human plasma for 10 min. 1test PM-2K Anti-Macrophage antibody (Abcam ab58822) was added to each 106 cells and incubated at 4 ℃ for 30 min. 5ml of PBS was added, 400g was centrifuged for 5min, and the supernatant was discarded. Resuspend cells and add 1test Goat Anti-Mouse lgG Fc, Human/Bovine/Horse SPads-FTC (southern Biotech 1013-02) per 106 cells and incubate for 30min at 4 ℃. Resuspend antibody add 5ml PBS, 400g centrifugation for 5min, discard supernatant, repeat the previous step. The cells were resuspended and 1test PerCP anti-human CD3(Biolegend 300427), 1test PE anti-human CD19(Biolegend 302207), 1test APCanti-human CD14(Biolegend 325608) were added to 106 cells and incubated at 4 ℃ for 30 min. Another tube of cells was added with isotype control as above. And (4) washing away the unbound antibody of the incubated sample, and performing flow cytometry detection. And (3) setting a gate to circle out a cell population of CD3-CD19-, and detecting the macrophage levels of the PM-2K + CD14+ and the PM-2K + CD 14-.

1.3.2 detection of peripheral blood IL-10: 5ml of anticoagulated whole blood is extracted, placed in an EDTA anticoagulation tube and kept stand for 1h, 3600 Xg is carried out, the serum is separated by centrifugation for 10min and stored at-80 ℃ for standby. And (3) carrying out quantitative detection on IL-10 in serum by using an enzyme-linked immunosorbent assay (ELISA), wherein the operation steps are strictly carried out according to the kit instructions.

1.4 methods of treatment

All patients entered the group as 1: 1 ratio was randomly divided into 2 groups, one group was the astragalus prescription group, the other group was the placebo group:

the method comprises the following steps: 30g of astragalus, 30g of curcuma zedoary, 15g of Chinese sage herb, 15g of codonopsis pilosula and 9g of medlar. The preparation is made into granules by Jiangyin Tianjiang pharmaceutical industry, Inc. of Chinese medicine group, 1 dose per day, and is taken with boiled water for 2 times.

② placebo: the simulated traditional Chinese medicine compound preparation particles are prepared from auxiliary materials and flavoring agents, and the package, the appearance, the weight, the smell and the color of the simulated traditional Chinese medicine compound preparation particles are consistent with those of the traditional Chinese medicine preparation. The preparation is made into granules by Jiangyin Tianjiang pharmaceutical industry, Inc. of Chinese medicine group, 1 dose per day, and 2 times for administration.

1.5 Observation index

1.5.1 Main Observation indicators

Changes in the expression of IL-10, CD14+ macrophages in the peripheral blood of two groups of breast cancer patients before and after treatment were recorded. According to the constructed IL-10 and CD14+ macrophage combined detection formula: and y is an IL-10 value +3.62 CD14+ macrophage value, and the obtained combined assay data y is calculated.

1.5.2 Secondary Observation indicators

(1) Quality of life scoring: through questionnaire survey of a quality of life scale (QLQ-C30), the score RS of each field can be calculated by adding the item scores included in each field of the scale and dividing the number of the items included, namely the RS ═ Q1+ Q2+. + Qn)/n, and then the RS is converted into a standardized score SS with the value between 0 and 100. The scoring rules specify: a higher score for the overall health status and functional domain indicates better functional status or quality of life, while a higher score for the symptom domain indicates more symptoms and problems, worse quality of life,

(2) evaluation of the curative effect of the traditional Chinese medicine symptoms: grading according to the degree of severity of symptoms, and according to the principal symptoms: no symptom 0 point; mild grade 2 points; medium 4 points; the gravity was 6 points. The secondary symptoms are as follows: no symptom 0 point; mild degree 1 score; medium 2 points; and 3, observing and recording before and after treatment, calculating symptom integral, and evaluating the curative effect according to the integral. The formula (nimodipine method) is calculated as follows:

the curative effect score n is [ (total number of syndrome integrals before treatment-total number of syndrome integrals after treatment)/total number of syndrome integrals before treatment ] × 100%

The method is remarkably improved: after treatment, the syndrome is obviously reduced, and n reaches 70 percent or more;

secondly, partial improvement: the symptoms are reduced after treatment, n is more than or equal to 30% and less than 70%;

③ no improvement: no or slight decrease in the symptoms after treatment, and n is only below 30%.

(3) The safety index is as follows: safety evaluation observation according to the evaluation Standard of clinical safety 4

Grade 1 is no adverse reaction;

grade 2 indicates mild adverse reaction, and the drug can be continuously administered without any treatment;

grade 3 indicates that the drug has moderate adverse reaction and can be continuously administered after being treated;

grade 4 indicates severe adverse reactions, and the test was withdrawn due to adverse reactions.

1.6 statistical methods

All data was entered into the computer, and the database was built using the SPSS 20.0 software package and analyzed. The measurement data is expressed by mean +/-standard deviation, and t test is used for conforming to normal distribution, and non-parameter test is used for not conforming to normal distribution. The comparison between groups of counted data was checked by t. And (5) carrying out rank sum check on the grade data. Pearson product difference correlation coefficients are selected for bivariate normal distribution data correlation analysis, and Spearman rank correlation coefficients are used when the application conditions of the product difference correlation analysis are not met. Linear trend testing of ordered grouped data using X ² And (6) checking. With P<0.05 indicates that the difference is statistically significant.

2 results

2.1 comparison of expression levels of IL-10 and CD14+ in peripheral blood between the Qishu group and the placebo group

By flow cytometry, we obtained the percentages of the peripheral blood CD14 macrophages before and after intervention in the stilbene surgery and placebo groups (see figures 1-A, B, C, D). The baseline IL-10 and CD14+ macrophages tested in combination prior to intervention showed no statistical differences (P > 0.05). After the traditional Chinese medicine is taken orally for 14 days, the IL-10 and CD14+ macrophages of patients in the astragalus prescription group are combined to detect that the expression displacement is 30.12 +/-6.57, and the expression level is obviously lower than that of a placebo group (P is 0.001) compared with that before intervention (P is less than 0.001). There was no significant change in IL-10 expression (P >0.05) when tested in combination with CD14+ macrophages before and after the placebo intervention (see Table 4, FIGS. 3-E, F).

TABLE 4 comparison of the stilbene operating prescription and placebo groups between groups in which expression was detected in combination with peripheral blood IL-10 and CD14+

2.2 comparison of the Effect of the stilbene procedure on the expression of the Combined detection of IL-10 and CD14+ in peripheral blood of patients with different clinical pathological characteristics

Through traditional Chinese medicine intervention, the IL-10 and CD14+ macrophage of II + III stage patients are detected to be combined to reduce the expression to 30.43 +/-6.65, the expression is obviously reduced (P is 0.018), and the changes of I stage patients before and after intervention are not statistically different (P is more than 0.05); the expression of IL-10 and CD14+ macrophages is obviously reduced (P is less than 0.05) when the combined detection is carried out on patients with different hormone receptor states and different HER2 expressions, but no obvious difference is found among the groups; the expression of IL-10 and CD14+ combined detection of all molecular profiles showed a decrease trend, but the decrease of the expression of IL-10 and CD14+ combined detection of only Luminal type B patients was statistically significant (P < 0.05). (see Table 5)

TABLE 5 stilbene therapy groups of peripheral blood IL-10 and CD14+ for different clinical and pathological characteristics

2.3 comparison of the quality of Life Scoring changes between the Astragalus prescription group and the placebo group

The higher the score of the functional scale in the quality of life score, the better the quality of life. Before treatment, the scores between the two groups were not statistically different before intervention (P >0.05), and were comparable, wherein the patients in the stilbene formula group and the placebo group both had emotional dysfunction such as stress, anxiety, and the like, and therefore the scores were relatively low, the scores were 39.79 + -21.80 in the stilbene formula group and 34.17 + -24.52 in the placebo group. After the traditional Chinese medicine is taken orally for 14 days, the emotional function score of the stilbene-art prescription composition is obviously improved (P is less than 0.05); no significant change was seen in the remaining functional scores (P > 0.05). (see Table 6)

TABLE 6 comparison of survival Scale function scores for the Astragalus surgery formula group and placebo group

The higher the score of the symptom scale in the quality of life score, the worse the quality of life. The scores between the two groups before treatment were not statistically different (P >0.05) and were comparable; after the traditional Chinese medicine is taken orally for 14 days, the fatigue, insomnia and overall health condition of the astragalus prescription group are obviously improved (P is less than 0.05), and no obvious difference is seen between the group and the group after the placebo group has a dry prognosis (P is more than 0.05). (see Table 7)

TABLE 7 comparison of symptom function scores for the stilbene technical formula group and the placebo group on the survival Scale

2.4 comparison of the improvement of the symptoms of the traditional Chinese medicine between the Qishu prescription group and the placebo group

Before treatment, the scores between the two groups were not statistically different (P >0.05) and were comparable. After the prognosis of the Chinese medicine dryness, the symptoms of weakness, lassitude, hypologia, sallow or dull complexion and spontaneous perspiration and night sweat in the secondary symptoms and the total scoring condition of the stilbene prescription group are obviously improved (P is less than 0.05), while no obvious difference is seen between the groups and the placebo group after the prognosis of the Chinese medicine dryness (P is more than 0.05), which are shown in tables 8 and 9.

TABLE 8 comparison of Chinese medicine symptom Master scores between the Astragalus membranaceus prescription group and placebo group

TABLE 9 comparison of the secondary symptom scores of the traditional Chinese medicine with the placebo group

The curative effect evaluation shows that the effective rate of the astragalus mongholicus formula is 40%, the effective rate of the placebo group is 7.5%, and the curative effect distribution of the two groups is also statistically different (P is less than 0.05), which is shown in table 10.

TABLE 10 evaluation of the improvement of the symptoms of Chinese medicine in the stilbene operation prescription group and placebo group

Discussion of 3

TME is important soil for the occurrence and development of breast cancer. The interaction of TME with different cell populations determines the biological behavior of the tumor. Macrophages are one of the most abundant cell populations in TME for breast cancer, accounting for approximately more than 50% of the number of immune cells, and their polarization of M2-type macrophages also suggests a worse prognosis outcome for breast cancer. In the early stage of the invention, M2 type macrophages are used as a starting point for exploration, and the results prove that the combined detection of IL-10 and CD14+ macrophages can reflect the tumor load and the tumor cell proliferation and differentiation capacity of a breast cancer patient, and has a certain prompting effect on the internal environment of tumor immunity and the prognosis outcome of breast cancer, so that the stilbene operation formula intervention is carried out on the breast cancer perioperative patient, the change of the combined detection expression level of IL-10 and CD14+ macrophages is observed, and the combined detection expression level of IL-10 and CD14+ macrophages in the stilbene operation formula group is obviously lower than that in a placebo group (P is less than 0.05) after the dry prognosis of traditional Chinese medicines. In the further stratification analysis of the stilbene-technique prescription group, the intervention effect of the stilbene-technique prescription on patients with terminal and hormone receptor negativity is also very obvious (P < 0.05). Proves that the astragalus operating method can influence the invasion of tumor cells and regulate and control the immune microenvironment of breast cancer tumors by regulating the expression of peripheral blood macrophages.

It is worth mentioning that the reduction in expression of the Luminal type B patient CD14+ macrophages in combination with the assay was most significant in the four molecular profiles (P < 0.05). Luminal type B is often characterized by low or no expression of ER, low or no expression of PR, insensitive endocrine treatment, more aggressive proliferation rate, prominent Gene mutation rate, and resistance to endocrine secretion, which limit the benefit of Luminal type B patients in Breast Cancer treatment (Zhu YY, Zhuao YC, Chen C, et al. CCL5 secreted by B Breast Cancer cells and administration of M2 secreted activities of MEK/STAT3 Breast Cancer CCR5[ J ] Gene,2022,812:146100.Tran B, Bedard. Luminal-B cultured and novel therapeutic targets [ J ] Breast, Cancer, 2011,13(6): 221). Furthermore, Luminal B-type has also been shown to express M2-type Macrophages with high TME levels, which may contribute to the internal environment conducive to tumor survival, leading to metastatic invasion (Raschini C, Bottai G, Sagona A, et al CXCR4/CXCL12 signalling and Protomor Macrophages in Primary Tumors and Sentinel Lymph node area invent in Luminal B Breast Cancer progress [ J ]. Dis Markers,2018,2018:5018671.), whereas the present invention suggests that stilbene-type has a more prominent regulatory role in peripheral blood M2-type Macrophages of Luminal B-type Breast Cancer patients, and may become an important breakthrough in Luminal B-type Breast Cancer treatment bottlenecks.

In addition, the quality of life scoring results show that both groups of patients have a certain degree of emotional management disorder, which is mainly manifested as anxiety, tension, low mood and the like; the symptom scale shows that fatigue and insomnia are commonly existed. Breast diseases are indistinguishable from emotion, "depression hurts the liver, anxiety hurts the spleen", depression and uneasiness, liver qi is accumulated, stagnation turns into fire, liver fire disturbs spirit, and heart spirit is disordered, which finally causes insomnia, insomnia and restlessness; excessive thinking can lead to spleen qi stagnation, deficient earth, poor source of qi and blood, malnutrition of the meridians and collaterals, and impairment of healthy qi, which can ultimately lead to consumptive disease and debilitation, and the deficiency can be cured, all of which are caused by the liver and spleen. In addition, the patients with breast cancer have serious qi consumption and blood injury in the perioperative period, damaged veins, easy influence on the acquired root, and more obvious symptoms of qi-blood stagnation and liver-spleen disorder. In the formula of the astragalus and the rhizoma atractylodis, the astragalus and the radix codonopsis tonify qi of the liver and the spleen, the rhizoma zedoariae and the Chinese sage penetrate through the Chinese wolfberry to dispel stasis of the liver and the spleen, and the medlar is cooperated to nourish deficiency and essence deficiency, so that the spleen and the earth are in good circulation, the liver and the collaterals are smooth, and the effects of regulating the liver and the spleen and harmonizing qi and blood are achieved. Therefore, after the prognosis of the Chinese medicine, the symptoms of anxiety, fatigue and insomnia of the stilbene prescription group are obviously relieved.

The invention discovers that the syndrome of qi deficiency and blood stasis is the main syndrome type of the patient in the perioperative period of the breast cancer, and the evaluation result of the clinical symptoms of the traditional Chinese medicine discovers that the postoperative patient has qi consumption and essence injury, has weak and lassitude, can also gather in the operative wound due to water dampness, has heat transformation due to depression, has recklessly blood flow due to blood heat, floats on the skin and has the red and swollen petechia petechiae; spleen qi deficiency failing to consolidate body fluids, liver fire stagnates and evaporates, causing disharmony between ying and wei, resulting in spontaneous sweating and night sweat. The qi shu formula strengthens body resistance and removes blood stasis, and the formula is warm but not hot, and can nourish yin and fluid and harmonize yingfen and weifen. The clinical symptoms of qi deficiency and blood stasis in traditional Chinese medicine can be obviously improved by applying the astragalus root technique to the symptoms.

In conclusion, the stilbene operation method can obviously reduce the combined detection expression of IL-10 and CD14+ macrophages in the peripheral blood of a breast cancer patient, and the stilbene operation method is changed from a middle-advanced patient and a Luminal B-type patient to a later-advanced patient, so that the stilbene operation method is prompted to possibly intervene in the expression of M2 macrophages in the internal environment of the body of the breast cancer patient to regulate and control TME so as to improve prognosis; the astragalus root operation prescription has the advantages of tonifying qi of the liver and the spleen, breaking stasis of the liver and the spleen, strengthening spleen and activating spleen, smoothing liver collaterals, achieving the effects of regulating the liver and the spleen and nourishing qi and blood, being favorable for regulating emotion management disorder caused by emotional disorder after operation of a breast cancer patient, improving symptoms of qi deficiency and blood stasis such as qi deficiency fatigue, insomnia, spontaneous perspiration and the like, and improving the life quality of the patient.

While the preferred embodiments of the present invention have been described in detail, it will be understood by those skilled in the art that the invention is not limited thereto, and that various changes and modifications may be made without departing from the spirit of the invention, and the scope of the appended claims is to be accorded the full range of equivalents.

Claims

1. A traditional Chinese medicine composition for improving immune microenvironment and quality of life in a breast cancer perioperative period is characterized by being prepared from the following raw material medicines in parts by weight: 15-45 parts of astragalus root, 15-45 parts of zedoary, 5-25 parts of Chinese sage herb, 5-25 parts of codonopsis pilosula and 3-18 parts of medlar.

2. The traditional Chinese medicine composition according to claim 1, which is prepared from the following raw material medicines in parts by weight: 20-40 parts of astragalus membranaceus, 20-40 parts of curcuma zedoary, 10-20 parts of Chinese sage herb, 10-20 parts of codonopsis pilosula and 6-12 parts of medlar.

3. The traditional Chinese medicine composition according to claim 1, which is prepared from the following raw material medicines in parts by weight: 30 parts of astragalus, 30 parts of curcuma zedoary, 15 parts of Chinese sage herb, 15 parts of codonopsis pilosula and 9 parts of medlar.

4. Use of the Chinese medicinal composition of any one of claims 1-3 in the preparation of a medicament for improving the microenvironment and quality of life of qi deficiency and blood stasis type breast cancer in the perioperative period.

5. A pharmaceutical preparation using the traditional Chinese medicine composition for improving qi deficiency and blood stasis type breast cancer perioperative immune microenvironment and quality of life as an active ingredient as claimed in any one of claims 1 to 3, wherein the pharmaceutical preparation is prepared into a pharmaceutically common dosage form by adopting a conventional preparation method in the field.

6. The Chinese medicinal composition according to claim 5, wherein the medicinal preparation is decoction, pill, tablet, mixture, capsule, granule, powder or paste.

7. The Chinese medicinal composition according to claim 6, wherein the medicinal preparation further comprises pharmaceutically acceptable adjuvants.