A Danshu Fuyuan Prescription, its preparation method and the application of the Danshu Fuyuan Prescription in the preparation of drugs for treating the syndrome of intermingled phlegm and stasis

Abstract

The present invention belongs to the field of pharmaceuticals, and particularly relates to a Danshu Fuyuan formula, a preparation method thereof, and the application of the Danshu Fuyuan formula in the preparation of a medicament for treating the syndrome of intermingled phlegm and stasis. The Danshu Fuyuan formula comprises the following raw materials in parts by weight: 25-35 parts of Salvia miltiorrhiza, 10-15 parts of Atractylodes macrocephala, 25-35 parts of Astragalus membranaceus, 10-20 parts of Lonicera japonica, 10-20 parts of Polygonum cuspidatum, 10-20 parts of Curcuma aromatica, 10-15 parts of Poria cocos, 25-35 parts of Coix lacryma-jobi, and 10-15 parts of Notopterygium incisum. Research shows that the Danshu Fuyuan formula of the present invention can effectively treat the syndrome of intermingled phlegm and stasis in coronary heart disease, the syndrome of intermingled phlegm and stasis in idiopathic membranous nephropathy, and the syndrome of intermingled phlegm and stasis in vascular dementia, providing a new idea for the treatment of the syndrome of intermingled phlegm and stasis.

Description

Technical Field

[0001] The present invention belongs to the field of medicines, and particularly relates to a Danshu Fuyuan formula, a preparation method thereof, and an application of the Danshu Fuyuan formula in the preparation of a medicine for treating the syndrome of intermingled phlegm and stasis. Background Art

[0002] The syndrome of intermingled phlegm and stasis refers to: the intermingling of phlegm turbidity and stasis blood, with common symptoms such as local mass with stabbing pain, or limb numbness, flaccidity and paralysis, chest tightness with excessive phlegm, or phlegm with dark purple blood clots, purple and dark tongue or with spots, greasy tongue coating, and string-taut and unsmooth pulse.

[0003] Existing medicines can only treat a certain type of syndrome of intermingled phlegm and stasis during treatment. For example, a patent with the publication number CN116650602A discloses a traditional Chinese medicine composition, which is: 19 - 31 parts of Clematis chinensis, 24 - 36 parts of Chaenomeles sinensis, 24 - 36 parts of Astragalus membranaceus, 19 - 31 parts of Dipsacus asperoides, 19 - 31 parts of Taxillus chinensis, 15 - 27 parts of Paeonia lactiflora, 15 - 27 parts of Paeonia suffruticosa, 14 - 26 parts of Prunus mume, 9 - 21 parts of Vigna umbellata, 6 - 18 parts of Alpinia officinarum, and 4 - 16 parts of Glycyrrhiza uralensis. This composition is only applicable to the treatment of the syndrome of intermingled phlegm and stasis in femoral head necrosis, and the treatment scope is relatively narrow.

[0004] Therefore, based on this, the technical solution of the present invention is proposed. Summary of the Invention

[0005] In order to solve the problems existing in the prior art, the present invention provides a Danshu Fuyuan formula, which includes the following raw materials in parts by weight: 25 - 35 parts of Salvia miltiorrhiza, 10 - 15 parts of Atractylodes macrocephala, 25 - 35 parts of Astragalus membranaceus, 10 - 20 parts of Lonicera japonica, 10 - 20 parts of Polygonum cuspidatum, 10 - 20 parts of Curcuma aromatica, 10 - 15 parts of Poria cocos, 25 - 35 parts of Coix lacryma-jobi, and 10 - 15 parts of Notopterygium incisum.

[0006] In the formula, Salvia miltiorrhiza is bitter and slightly cold in nature, enters the pericardium and liver meridians, has the same effect as the four drugs of Siwu Decoction, and has the effect of cooling blood and promoting blood circulation, and is the monarch drug of this formula; Atractylodes macrocephala and Astragalus membranaceus are used as ministers to supplement vital energy. Lonicera japonica and Polygonum cuspidatum clear the heat in the blood vessels and disperse the formed nodules; Poria cocos and Coix lacryma-jobi are used as assistants to strengthen the spleen and resolve phlegm, and also make Astragalus membranaceus and Atractylodes macrocephala tonify without causing stasis; Curcuma aromatica combined with Lonicera japonica and Polygonum cuspidatum helps Salvia miltiorrhiza clear heat, cool blood and dissipate nodules; Notopterygium incisum is used as an assistant envoy. On the one hand, by virtue of its pungent and warm nature, the whole formula is not overly cold. On the other hand, with the ascending and clearing power of the wind drug, it promotes the rising of yang qi, and combined with Astragalus membranaceus and Atractylodes macrocephala, it restores the ascending and descending of the qi movement in the spleen and stomach. The whole formula combines cold and heat, combines attacking and tonifying, and has a scientific and reasonable compatibility.

[0007] Preferably, the Danshu Fuyuan formula includes the following raw materials in parts by weight: 30 parts of Salvia miltiorrhiza, 12 parts of Atractylodes macrocephala, 30 parts of Astragalus membranaceus, 15 parts of Lonicera japonica, 15 parts of Polygonum cuspidatum, 15 parts of Curcuma aromatica, 12 parts of Poria cocos, 30 parts of Coix lacryma-jobi, and 12 parts of Notopterygium incisum.

[0008] Based on the same inventive concept, the solution of the present invention is to provide a preparation method of Danshu Fuyuan Prescription, and the preparation method is as follows:

[0009] Crush the raw materials of Danshu Fuyuan Prescription and then mix them to obtain a composition;

[0010] Or, mix the raw materials of Danshu Fuyuan Prescription and then crush them to obtain a composition;

[0011] Or, extract and refine the raw materials of Danshu Fuyuan Prescription according to conventional processes to obtain one of tablets, capsules, granules, pills, powders or oral liquids.

[0012] Based on the same inventive concept, another solution of the present invention is to provide an application of Danshu Fuyuan Prescription in the preparation of drugs for treating phlegm stasis syndrome, and the phlegm stasis syndrome is one of coronary heart disease phlegm stasis syndrome, vascular dementia phlegm stasis syndrome, and idiopathic membranous nephropathy phlegm stasis syndrome.

[0013] Preferably, the drug is a drug for improving the cardiac function of coronary heart disease;

[0014] And / or, the drug is a drug for improving the myocardial ischemia state of coronary heart disease;

[0015] And / or, the drug is a drug for improving the pathological state of coronary heart disease;

[0016] And / or, the drug is a drug for improving the blood lipid level of coronary heart disease.

[0017] Preferably, the drug is a drug for improving the serum inflammation level of coronary heart disease;

[0018] And / or, the drug is a drug for improving the mRNA transcription of myocardial tissue of coronary heart disease;

[0019] And / or, the drug is a drug for improving the expression of inflammatory proteins in myocardial tissue of coronary heart disease;

[0020] And / or, the drug is a drug for improving the expression of pathway proteins in myocardial tissue of coronary heart disease.

[0021] Preferably, the drug is a drug for reducing the urine protein level.

[0022] Preferably, the drug is a drug for reducing the TCM syndrome score of patients with vascular dementia;

[0023] And / or, the drug is a drug for increasing the MMSE score of patients with vascular dementia;

[0024] And / or, the drug is a drug for increasing the MoCA score of patients with vascular dementia;

[0025] And / or, the drug is a drug for reducing the ADL score of patients with vascular dementia.

[0026] The beneficial effects of the present invention are as follows:

[0027] Research shows that the Danshu Fuyuan Prescription of the present invention can effectively treat the syndrome of phlegm and stasis obstruction in coronary heart disease, idiopathic membranous nephropathy, and vascular dementia, providing a new method for treating the syndrome of phlegm and stasis obstruction. Specifically:

[0028] (1) The Danshu Fuyuan Prescription of the present invention can regulate the levels of four blood lipids in rats with coronary heart disease, and the high-dose Danshu Fuyuan Prescription has a stronger ability to regulate blood lipids. In addition, the Danshu Fuyuan Prescription can inhibit the secretion of serum inflammatory factors and the expression of inflammatory proteins. And the Danshu Fuyuan Prescription may play a therapeutic role in rats with coronary heart disease by inhibiting the TLR4 / NF-κB pathway.

[0029] (2) The Danshu Fuyuan Prescription of the present invention can effectively reduce the 24-hour urinary protein level in patients with IMN, and the 24-hour urinary protein level shows an obvious downward trend with the prolongation of the treatment course. Further, according to the clinical efficacy criteria for IMN, the total remission rate of patients treated with the Danshu Fuyuan Prescription for 3 months is 37%, the total remission rate of patients treated for 6 months is 50%, and the total remission rate of patients treated for 12 months reaches 72%. And with the prolongation of the treatment course, the remission rate increases significantly.

[0030] (3) The Danshu Fuyuan Prescription of the present invention can reduce the TCM syndrome scores of VD patients, significantly improve the related TCM syndromes of VD patients; and can effectively improve the MMSE and MoCA scores of VD patients, significantly improving the cognitive function of patients; at the same time, it can effectively reduce the ADL score of VD patients and improve the daily living ability of VD patients. Brief Description of the Drawings

[0031] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0032] Figure 1 It is the detection result chart of serum CK-MB of rats in each group.

[0033] Figure 2 It is the detection result chart of serum LDH activity of rats in each group.

[0034] Figure 3 It is the planar detection chart of echocardiography of rats in the sham operation group.

[0035] Figure 4 It is the planar detection chart of echocardiography of rats in the model group.

[0036] Figure 5 It is the planar graph of cardiac ultrasound examination of rats in the low-dose group.

[0037] Figure 6 It is the planar graph of cardiac ultrasound examination of rats in the medium-dose group.

[0038] Figure 7 It is the planar graph of cardiac ultrasound examination of rats in the high-dose group.

[0039] Figure 8 It is the planar graph of cardiac ultrasound examination of rats in the positive drug group.

[0040] Figure 9 It is the detection result graph of EF of rats in each group.

[0041] Figure 10 It is the detection result graph of FS of rats in each group.

[0042] Figure 11 It is the detection result graph of IVSd of rats in each group.

[0043] Figure 12 It is the detection result graph of IVSs of rats in each group.

[0044] Figure 13 It is the anatomical graph of myocardial infarction of rats in each group.

[0045] Figure 14 It is the graph of myocardial infarction ratio (area) of rats in each group.

[0046] Figure 15 It is the HE staining result graph of myocardial tissue of sham-operated rats (×40).

[0047] Figure 16 It is the HE staining result graph of myocardial tissue of model rats (×40).

[0048] Figure 17 It is the HE staining result graph of myocardial tissue of rats in the low-dose group (×40).

[0049] Figure 18 It is the HE staining result graph of myocardial tissue of rats in the medium-dose group (×40).

[0050] Figure 19 It is the HE staining result graph of myocardial tissue of rats in the high-dose group (×40).

[0051] Figure 20 It is the HE staining result graph of myocardial tissue of rats in the positive drug group (×40).

[0052] Figure 21 It is the Masson staining result graph of myocardial tissue of sham-operated rats (×40).

[0053] Figure 22 It is the Masson staining result diagram of the myocardial tissue of the model group rats (×40).

[0054] Figure 23 It is the Masson staining result diagram of the myocardial tissue of the low-dose group rats (×40).

[0055] Figure 24 It is the Masson staining result diagram of the myocardial tissue of the medium-dose group rats (×40).

[0056] Figure 25 It is the Masson staining result diagram of the myocardial tissue of the high-dose group rats (×40).

[0057] Figure 26 It is the Masson staining result diagram of the myocardial tissue of the positive drug group rats (×40).

[0058] Figure 27 It is the detection result diagram of the TC content of rats in each group.

[0059] Figure 28 It is the detection result diagram of the TG content of rats in each group.

[0060] Figure 29 It is the detection result diagram of the LDL-C content of rats in each group.

[0061] Figure 30 It is the detection result diagram of the HDL-C content of rats in each group.

[0062] Figure 31 It is the detection result diagram of the IL-1β content of rats in each group.

[0063] Figure 32 It is the detection result diagram of the IL-6 content of rats in each group.

[0064] Figure 33 It is the detection result diagram of the TNF-α content of rats in each group.

[0065] Figure 34 It is the detection result diagram of the MPO content of rats in each group.

[0066] Figure 35 It is the detection result diagram of the Resistin content of rats in each group.

[0067] Figure 36 It is the detection result diagram of the iNOS content of rats in each group.

[0068] Figure 37 It is the detection result diagram of the MMP-8 content of rats in each group.

[0069] Figure 38It is the detection result graph of the MMP-9 content in each group of rats.

[0070] Figure 39 It is the detection result graph of the ACE content in each group of rats.

[0071] Figure 40 It is the detection result graph of the IL-6 mRNA in each group of rats.

[0072] Figure 41 It is the detection result graph of the TNF-α mRNA in each group of rats.

[0073] Figure 42 It is the detection result graph of the MCP-1 mRNA in each group of rats.

[0074] Figure 43 It is the detection result graph of the LP-PLA2 mRNA in each group of rats.

[0075] Figure 44 It is the change graph of the inflammatory protein expression level in the myocardial tissue of each group of rats.

[0076] Figure 45 It is the detection result graph of NOD-2 in each group of rats.

[0077] Figure 46 It is the detection result graph of IL-1β in each group of rats.

[0078] Figure 47 It is the detection result graph of E-selectin in each group of rats.

[0079] Figure 48 It is the detection result graph of LOX-1 in each group of rats.

[0080] Figure 49 It is the detection result graph of CHI3L-1 in each group of rats.

[0081] Figure 50 It is the detection result graph of TIMP-3 in each group of rats.

[0082] Figure 51 It is the detection result graph of ACE-2 in each group of rats.

[0083] Figure 52 It is the detection result graph of GDF-15 in each group of rats.

[0084] Figure 53 It is the change of the expression level of the pathway protein in the myocardial tissue of each group of rats.

[0085] Figure 54 It is the detection result graph of TLR4 in each group of rats.

[0086] Figure 55It is the detection result diagram of cytoplasmic NF-κB P65 in each group of rats.

[0087] Figure 56 It is the detection result diagram of nuclear NF-κB P65 in each group of rats. Specific implementation mode

[0088] To make the objectives, technical solutions and advantages of the present invention clearer, the technical solutions of the present invention will be described in detail below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other implementation manners obtained by those of ordinary skill in the art based on the embodiments in the present invention without creative efforts belong to the scope protected by the present invention.

[0089] Example 1

[0090] This example provides a preparation method of Danshu Fuyuan Prescription, specifically: 25 g of Salvia miltiorrhiza, 10 g of Atractylodes macrocephala, 25 g of Astragalus membranaceus, 10 g of Lonicera japonica, 10 g of Polygonum cuspidatum, 10 g of Curcuma aromatica, 10 g of Poria cocos, 25 g of Coix lacryma-jobi and 10 g of Notopterygium incisum are pulverized and then mixed to obtain a composition, and then it is conventionally processed into tablets.

[0091] Example 2

[0092] This example provides a preparation method of Danshu Fuyuan Prescription, specifically: 35 g of Salvia miltiorrhiza, 15 g of Atractylodes macrocephala, 35 g of Astragalus membranaceus, 20 g of Lonicera japonica, 20 g of Polygonum cuspidatum, 20 g of Curcuma aromatica, 15 g of Poria cocos, 35 g of Coix lacryma-jobi and 15 g of Notopterygium incisum are mixed and then pulverized to obtain a composition, and then it is conventionally processed into capsules.

[0093] Example 3

[0094] This example provides a preparation method of Danshu Fuyuan Prescription, specifically: 30 g of Salvia miltiorrhiza, 12 g of Atractylodes macrocephala, 30 g of Astragalus membranaceus, 15 g of Lonicera japonica, 15 g of Polygonum cuspidatum, 15 g of Curcuma aromatica, 12 g of Poria cocos, 30 g of Coix lacryma-jobi and 12 g of Notopterygium incisum are mixed, and then extracted and refined according to the conventional process to obtain granules.

[0095] Example 4

[0096] The difference between Example 4 and Example 3 is that Danshu Fuyuan Prescription is made into pills.

[0097] Example 5

[0098] The difference between Example 5 and Example 3 is that Danshu Fuyuan Prescription is made into powders.

[0099] Example 6

[0100] The difference between Example 6 and Example 3 is that Danshu Fuyuan Prescription is made into oral liquids.

[0101] Application Example 1: Application of Danshu Fuyuan Prescription in the Preparation of Drugs for Treating Coronary Heart Disease with Syndrome of Phlegm-Stasis Obstruction (Using the Granules Obtained in Example 3 as the Drug)

[0102] (I) Experimental Method

[0103] Eighty-five SPF male Wistar rats, weighing 180±20 g, were randomly divided into a sham operation group, a model group, a low-dose Danshu Fuyuan Prescription group (prepared in Example 3, hereinafter referred to as the low-dose group), a medium-dose Danshu Fuyuan Prescription group (prepared in Example 3, hereinafter referred to as the medium-dose group), a high-dose Danshu Fuyuan Prescription group (prepared in Example 3, hereinafter referred to as the high-dose group), and a positive drug group (the positive drug is Danlou Tablets). Among them, there were 10 rats in the sham operation group, and 15 rats in each of the other groups.

[0104] The sham operation group was fed with ordinary feed every day, and the other groups of rats (75 rats) were fed with high-fat feed every day for 8 consecutive weeks. Four weeks after feeding with high-fat feed, the left anterior descending branch of the coronary artery of the rats in the model group was ligated for modeling. On the second day after the operation, the rats in the positive drug group (left anterior descending branch ligation of the coronary artery + Danlou Tablets) were given 0.47 g / Kg aqueous solution of Danlou Tablets by gavage, and the rats in the low-dose group, medium-dose group, and high-dose group (left anterior descending branch ligation of the coronary artery + Danshu Fuyuan Prescription) were given 1.25 g / Kg, 2.5 g / Kg, and 5 g / Kg aqueous solution of Danshu Fuyuan Prescription by gavage respectively. The rats in the coronary heart disease model group (left anterior descending branch ligation of the coronary artery) and the sham operation group (only threading without ligation) were given an equal amount of distilled water by gavage once a day for 4 consecutive weeks. Four weeks after administration, blood was taken from the abdominal aorta of the rats to measure the levels of serum total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C), and the levels of serum IL-6, TNF-α, IL-1β, MMP-9, MPO, Resistin, ACE, MMP-8, and iNOS were detected by enzyme-linked immunosorbent assay (ELISA). The heart tissues of the rats were taken, and HE staining and Masson staining were used to observe the proliferation of myocardial cells and collagen. The mRNA expression levels of myocardial IL-6, TNF-α, MCP-1, and LP-PLA2 were detected by PCR, and the protein expressions of NOD-2, IL-1β, E-selection, LOX-1, CHI3L-1, TIMP-3, GDF-25, and ACE-2 in the myocardial tissue, as well as the protein levels of TLR4, cytoplasmic NF-κBp65, and nuclear NF-κBp65 were detected by Western Blot.

[0105] (II) Experimental Contents and Results

[0106] 1. Effect of Danshu Fuyuan Prescription on Myocardial Enzymes in Rats with Coronary Heart Disease:

[0107] After 4 weeks of intervention with Danshu Fuyuan Formula, the levels of serum myocardial enzymes in coronary heart disease rats were detected. The specific results are shown in Figure 1 , Figure 2 . Compared with the sham operation group, the activities of serum CK-MB and LDH in the model group were significantly increased (▲▲▲▲P<0.0001), showing statistical differences; compared with the model group, the activities of CK-MB and LDH in the low-dose group, medium-dose group, and high-dose group were significantly decreased (****P<0.0001).

[0108] The results showed that Danshu Fuyuan Formula could improve the levels of myocardial enzymes in coronary heart disease rats.

[0109] 2. Effects of Danshu Fuyuan Formula on cardiac function in coronary heart disease rats:

[0110] After 4 weeks of intervention with Danshu Fuyuan Formula, the cardiac function was detected by echocardiography. The specific results are shown in Figures 3 to 12 . Compared with the sham operation group, the EF and FS of rats in the model group were significantly decreased (▲▲▲▲P<0.0001), and the IVSd and IVSs were significantly thinned (▲▲P<0.01 or ▲▲▲▲P<0.0001), showing statistical differences. Compared with the model group, the low-dose group, medium-dose group, and high-dose group could significantly increase the levels of EF, FS, IVSd, and IVSs (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001), and showed a dose-dependent manner.

[0111] The results showed that Danshu Fuyuan Formula could improve the cardiac function in coronary heart disease rats.

[0112] 3. Effects of Danshu Fuyuan Formula on myocardial ischemia in coronary heart disease rats:

[0113] As Figure 13 shown, the normal myocardial tissue of rats presented dark red, while the infarcted myocardial tissue presented pale white. The infarct area was measured using Image J software. As the results in Figure 14 showed, compared with the sham operation group, the infarct area of rats in the model group was significantly increased (▲▲▲▲P<0.0001), showing statistical differences, indicating successful modeling. Compared with the model group, the infarct areas of rats in the low-dose group, medium-dose group, and high-dose group were significantly decreased (***P<0.001).

[0114] The results showed that Danshu Fuyuan Formula could improve the myocardial ischemia state in coronary heart disease rats.

[0115] 4. Effects of Danshu Fuyuan Formula on myocardial pathological state in coronary heart disease rats:

[0116] As Figures 15 to 20As shown, the HE staining results showed that the overall structure of the myocardial tissue in the sham operation group of rats was intact, the myocardial cells were of normal size, arranged neatly and densely, with almost no infiltration of inflammatory cells, and no atrophy or apoptosis of muscle fibers. Compared with the sham operation group, the myocardial cells in the model group of rats were arranged disorderly and loosely, the tissue was discontinuous, there was infiltration of inflammatory cells between cells, and obvious interstitial edema. Compared with the model group, after treatment with the low-dose group, medium-dose group, and high-dose group, the above conditions were alleviated to varying degrees, the arrangement of the myocardial tissue was basically regular and dense, and there was a small amount of infiltration of inflammatory cells.

[0117] Masson staining was used to analyze the degree of myocardial fibrosis in rats of each group. As Figures 21 to 26 shown, the area stained blue was the positive area of fibrosis. In the myocardial tissue of the sham operation group of rats, the myocardial cells were arranged neatly, the myocardial fibers were arranged regularly, and a small amount of stained fibrous tissue was visible in focal areas, with only a small amount of collagen fiber hyperplasia. In the model group of rats, a large number of myocardial cells were necrotic, the remaining myocardial cells were arranged disorderly, the myocardial cells were sparse, and myocardial fibrosis was obvious. The degree of myocardial fibrosis in the low-dose group, medium-dose group, and high-dose group of rats was significantly reduced, the fibrotic area was significantly reduced, and the drug effect was significant.

[0118] The results showed that Danshu Fuyuan Prescription could improve the pathological state of coronary heart disease rats.

[0119] 5. Effects of Danshu Fuyuan Prescription on blood lipids in coronary heart disease rats:

[0120] Compared with the sham operation group, the contents of serum TC, TG, and LDL-C in the model group of rats were significantly increased (▲▲▲P<0.001 or ▲▲▲▲P<0.0001), and the level of HDL-C was significantly decreased (▲▲▲P<0.001). Compared with the model group, the TC levels in the medium-dose group, high-dose group, and positive drug group were significantly decreased (*P<0.05, ****P<0.0001), the TG and LDL-C levels in the high-dose group and positive drug group were significantly decreased (**P<0.01), while the HDL-C level was significantly increased (*P<0.05). The specific results are shown in Figures 27 to 30 .

[0121] The results showed that Danshu Fuyuan Prescription could improve the blood lipid levels of coronary heart disease rats.

[0122] 6. Effects of Danshu Fuyuan Prescription on serum inflammatory factors in coronary heart disease rats:

[0123] The effects of Danshu Fuyuan Prescription on the levels of serum IL-1β, IL-6, TNF-α, MPO, Resistin, iNOS, MMP-8, MMP-9, and ACE in coronary heart disease rats are shown in Figures 31 to 39Compared with the sham operation group, the levels of IL-1β, IL-6, TNF-α, MPO, Resistin, iNOS, MMP-8, MMP-9, and ACE in the model group of rats were significantly increased (▲▲▲▲P<0.0001); compared with the model group of rats, the levels of IL-1β, IL-6, TNF-α, Resistin, MMP-9, and ACE in the low-dose group, medium-dose group, and high-dose group of rats were significantly decreased (*P<0.05, **P<0.01, ***P<0.001, or ****P<0.0001). The levels of MMP-8 in the medium-dose group and high-dose group of rats were significantly decreased (***P<0.001, ****P<0.0001). Only the levels of serum MPO and iNOS in the high-dose group of rats were significantly decreased (****P<0.0001).

[0124] The results showed that Danshu Fuyuan Formula could improve the serum inflammatory levels in rats with coronary heart disease.

[0125] 7. Changes in the mRNA levels of inflammatory factors in the myocardial tissue of rats with coronary heart disease treated with Danshu Fuyuan Formula:

[0126] Compared with the sham operation group, the transcriptional levels of IL-6, TNF-α, MCP-1, and LP-PLA2 mRNA in the myocardial tissue of the model group of rats were significantly increased (▲▲▲▲P<0.0001); compared with the model group, the transcriptional levels of IL-6, TNF-α, MCP-1, and LP-PLA2 mRNA in the myocardial tissue of the low-dose group, medium-dose group, and high-dose group of rats were decreased (****P<0.0001). The results are shown in Figures 40 to 43 .

[0127] The results showed that Danshu Fuyuan Formula could improve the mRNA transcription in the myocardial tissue of rats with coronary heart disease.

[0128] 8. Effects of Danshu Fuyuan Formula on the expression of inflammatory proteins in the myocardial tissue of rats with coronary heart disease:

[0129] The effects of Danshu Fuyuan Formula on the protein expression of NOD-2, IL-1β, E-selectin, LOX-1, CHI3L-1, TIMP-3, ACE-2, and GDF-15 in the myocardium of rats with coronary heart disease are shown in Figures 44 to 52Compared with the sham operation group, the protein levels of NOD-2, IL-1β, E-selectin, LOX-1, CHI3L-1, and GDF-15 in the myocardial tissues of rats in the model group were significantly increased (▲▲▲▲P<0.0001), while the expressions of TIMP-3 and ACE-2 were significantly decreased (▲▲▲▲P<0.0001). Compared with the model group, the levels of E-selectin, CHI3L-1, and GDF-15 in the myocardial tissues of rats in the low-dose group, medium-dose group, and high-dose group were significantly decreased (**P<0.01 or ****P<0.0001). The levels of NOD-2, IL-1β, and LOX-1 in the myocardial tissues of rats in the medium-dose group and high-dose group were significantly decreased, while the expressions of TIMP-3 and ACE-2 were significantly increased (**P<0.01 or ****P<0.0001).

[0130] The results showed that Danshu Fuyuan Formula could improve the expression of inflammatory proteins in the myocardial tissues of coronary heart disease rats.

[0131] 9. Effects of Danshu Fuyuan Formula on the expression of pathway proteins in the myocardial tissues of coronary heart disease rats:

[0132] Compared with the sham operation group, the expressions of TLR4 and nuclear NF-κB P65 in the myocardial tissues of rats in the model group were significantly increased (▲▲▲▲P<0.0001), while the cytoplasmic NF-κB P65 was significantly decreased. Compared with the model group, the expression of TLR4 in the myocardial tissues of rats in the medium-dose group and high-dose group was significantly decreased (****P<0.0001), the cytoplasmic NF-κB P65 was significantly increased (****P<0.0001), and the nuclear NF-κB P65 in the myocardial tissues of rats in the high-dose group was significantly decreased (****P<0.0001). The specific results are shown in Figures 53 to 56 。

[0133] The results showed that Danshu Fuyuan Formula could improve the expression of pathway proteins in the myocardial tissues of coronary heart disease rats.

[0134] Application Example 2: Application of Danshu Fuyuan Formula in the preparation of a drug for treating phlegm-stasis syndrome of idiopathic membranous nephropathy (IMN) (using the granule obtained in Example 3 as the drug)

[0135] (I) Research objects

[0136] 1. Source of cases: IMN subjects and healthy people who were treated in the outpatient clinics and wards of Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing Traditional Chinese Medicine Hospital, and Fangshan Traditional Chinese Medicine Hospital of Beijing from December 2019 to December 2021.

[0137] 2. Diagnostic criteria:

[0138] 2.1. Refer to "Nephrology (Third Edition)" edited by Wang Haiyan in 2008. The onset is insidious, often without a prodromal infection, and the medical history and physical examination both meet the diagnostic criteria for IMN.

[0139] 2.2. Refer to "Renal Biopsy Pathology (Third Edition)" edited by Zou Wanzhong in 2014. After renal tissue biopsy, it conforms to the light microscopy, electron microscopy, and immunofluorescence pathological characteristics of membranous nephropathy. Excluding secondary MN, the diagnosis is IMN, and it is staged according to the Ehrenreich-Churg staging method.

[0140] Stage I: The glomerular basement membrane is basically normal, with small and scattered electron-dense deposits irregularly distributed along the epithelial side of the basement membrane; Stage II: The deposits increase compared to Stage I, with the formation of spike projections, accompanied by extensive disappearance of epithelial cell foot processes; Stage III: The spike projections fuse, and the deposited material is irregular and sparse in texture, showing a worm-eaten appearance; Stage IV: The basement membrane is significantly irregularly thickened, and the density of the deposited electron-dense material decreases.

[0141] 2.3. Refer to "Research on the Macroscopic Diagnostic Criteria for the Phlegm-Stasis Syndrome of Coronary Heart Disease" published in "Chinese Journal of Integrated Traditional and Western Medicine" in 2016, and formulate the TCM diagnostic criteria for the phlegm-stasis syndrome of IMN, as shown in Table 1.

[0142] Table 1 Clinical Diagnostic Criteria for the Phlegm-Stasis Syndrome of Idiopathic Membranous Nephropathy

[0143]

[0144] (II) Research Plan

[0145] 1. Treatment and Diagnosis Plan:

[0146] 1.1. Basic Treatment: A low-salt, low-fat, and light diet, and use ACEI and ARB drugs within the tolerable range of the patient. Control the blood pressure within 130 / 80 mmHg. If the blood pressure has not reached the target blood pressure level (target blood pressure: systolic blood pressure ≤ 130 mmHg / and diastolic blood pressure ≤ 80 mmHg), CCB preparations, β-blockers, and diuretics can be used in combination.

[0147] 1.2. Clinical Trial Method: IMN Subject Group: Basic Treatment + Danshu Fuyuan Formula (Granules). Danshu Fuyuan Formula (Granules) is composed of Salvia miltiorrhiza, Atractylodes macrocephala, Curcuma aromatica, Coix lacryma-jobi, Astragalus membranaceus, Poria cocos, Polygonum cuspidatum, Lonicera japonica, and Notopterygium incisum. Efficacy: Tonifying qi, resolving phlegm, promoting blood circulation, clearing heat, and dissipating nodules. Dosage form: Granules. Take 1 dose of traditional Chinese medicine every day, divided into two times, morning and evening, for a course of 6 months.

[0148] 1.3. Combined Medication: For patients with combined hyperlipidemia, statin lipid-lowering drugs can be administered; for those with combined hypoproteinemia, plasma albumin lower than 25 g / L, and severe edema, human albumin can be intravenously infused; for those with hypercoagulable state and thrombosis risk, mai xuekang, aspirin, clopidogrel or low molecular weight heparin can be given; when accompanied by severe respiratory tract infection, urinary tract infection, or gastrointestinal infection, antibiotics can be administered. For mild cases of respiratory tract infection, qingre keli, jin hua qinggan keli, etc. can be used for treatment; for mild cases of gastrointestinal infection, berberine hydrochloride, bacillus licheniformis, etc. can be used for treatment; for mild cases of urinary tract infection, relinqing keli, longqing tablets, etc. can be used for treatment; for those with dry stools, biantong capsules can be added.

[0149] 2. Observation Indicators and Methods:

[0150] 2.1. Clinical Efficacy Observation Indicators: (1) Main Efficacy Indicators: 24-hour urinary protein quantification (24hUTP), albumin (ALB). (2) Secondary Efficacy Indicators: total serum protein (TP), blood urea nitrogen (BUN), serum creatinine (Scr), estimated glomerular filtration rate (eGFR), and eGFR is calculated using the CKD-EPI formula.

[0151] 2.2. Evaluation of Traditional Chinese Medicine Syndrome Scores: Quantify each clinical symptom of the phlegm-stasis mutual binding syndrome, including: abdominal fullness and distension, poor appetite, heaviness of the head and body, edema, obesity, dark color of the lips or gums, sallow or dull complexion, dry and rough skin, cyanosis of the extremities, sticky stools, lethargy, sticky mouth. According to the severity, it is divided into none, mild, moderate, and severe, scored 0, 2, 4, 6 respectively, and tongue and pulse are not included in the scoring. Mild means occasional symptoms / slight feeling, moderate means continuous symptoms / obvious feeling, and severe means affecting life. The total score of the traditional Chinese medicine syndrome score is the sum of the scores of each symptom, and the higher the score, the more severe the symptoms. As shown in Table 2 specifically.

[0152] Table 2 Syndrome Score Table for Idiopathic Membranous Nephropathy with Phlegm-Stasis Mutual Binding

[0153]

[0154]

[0155] 3. Efficacy Evaluation:

[0156] 3.1. Clinical Efficacy Evaluation Criteria:

[0157] Refer to the IMN clinical efficacy criteria formulated in the 2012 KDIGO Clinical Practice Guidelines for Glomerulonephritis:

[0158] (1) Complete Remission: Urinary protein quantification is less than 0.3 g / d, reaching the standard twice with an interval of at least 1 week, and plasma albumin and serum creatinine are within the normal range;

[0159] (2) Partial remission: Urinary protein quantification is less than 3.5 g / d, and it has decreased by more than 50% from the peak value; reaching the standard twice with an interval of at least 1 week, normal or improved plasma albumin, and stable serum creatinine.

[0160] (3) Ineffective: The above criteria are not met.

[0161] 3.2、Evaluation criteria for traditional Chinese medicine syndromes:

[0162] Refer to the "Guidelines for Clinical Research of New Traditional Chinese Medicines - Guidelines for Clinical Research of New Traditional Chinese Medicines in the Treatment of Chronic Nephritis" in the "Trial Guidelines for Clinical Research of New Traditional Chinese Medicines".

[0163] (1) Clinical cure: Traditional Chinese medicine clinical symptoms and signs disappear or basically disappear, and the syndrome score decreases by ≥ 95%.

[0164] (2) Marked effect: Traditional Chinese medicine clinical symptoms and signs are significantly improved, and the syndrome score decreases by ≥ 70%.

[0165] (3) Effective: Traditional Chinese medicine clinical symptoms and signs are all improved, and the syndrome score decreases by ≥ 30%.

[0166] (4) Ineffective: Traditional Chinese medicine clinical symptoms and signs are not significantly improved, or even worsened, and the syndrome score decreases by less than 30%.

[0167] Note: The calculation formula (Nimodipine method) is: [(pre-treatment score - post-treatment score) ÷ pre-treatment score] × 100%.

[0168] (III) Research results

[0169] 1. Case inclusion: A total of 10 healthy subjects were included in this example, and 65 IMN subjects from Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing Traditional Chinese Medicine Hospital, and Fangshan Traditional Chinese Medicine Hospital of Beijing were included. Among them, 6 were mis-included, and 5 were lost to follow-up at 3 months of treatment. Finally, 59 subjects were included in the FAS set for statistical analysis, 54 subjects were included in the PPS set for statistical analysis, and 59 were included in the SS for safety analysis.

[0170] 2. General information and clinical data:

[0171] 2.1 Gender and age: Among the 54 IMN subjects, 29 were male, accounting for 53.7%, and 25 were female, accounting for 46.3%. The male-to-female ratio was 1.16:1. Among the 10 healthy control subjects, 5 were male, accounting for 50%, and 5 were female, accounting for 50%. The male-to-female ratio was 1:1. The chi-square test showed no significant difference in gender between the two groups (P > 0.05). The age of the IMN group ranged from 20 to 77 years, with an average age of 53.07 ± 12.86 years. The age of the healthy control group ranged from 28 to 63 years, with an average age of 46.6 ± 10.53 years. The t-test showed no significant difference in age between the two groups (P > 0.05). The specific results are shown in Table 3.

[0172] Table 3 Comparison of basic data between the IMN group and the healthy control group (x±s)

[0173]

[0174] Note: When comparing the IMN group with the healthy group, P < 0.05 indicates a statistically significant difference.

[0175] 2.2 CKD staging: Among the 54 IMN subjects, 36 had CKD stage 1, accounting for 66.7%; 14 had CKD stage 2, accounting for 25.9%; and 4 had CKD stage 3, accounting for 7.4%.

[0176] 2.3 Risk assessment: Among the 54 IMN subjects, 26 were at low risk, accounting for 48.1%; 23 were at medium risk, accounting for 42.6%; and 5 were at high risk, accounting for 9.3%.

[0177] 2.4 Pathological staging: Among the 54 IMN subjects, 19 had stage I, 13 had stage I-II, 11 had stage II, 2 had stage II-III, and 9 had atypical membranous nephropathy.

[0178] 2.5 Among the 54 IMN subjects, 5 had proteinuria within the range of nephrotic syndrome, accounting for 9.3% of the total.

[0179] 2.6 Distribution of comorbidities: Among the 54 IMN subjects, 16 had hypertension, accounting for 29.6%; 8 had hyperlipidemia, accounting for 14.8%; 7 had diabetes, accounting for 13.0%; 3 had carotid and lower extremity atherosclerosis, accounting for 5.6%; 2 had inflammatory skin diseases such as psoriasis and eczema, accounting for 3.7%; and 1 had each of coronary heart disease, lacunar infarction, respiratory diseases, hypokalemia, prostatic hyperplasia, and osteonecrosis of the femoral head, accounting for 1.9%.

[0180] 3. FAS analysis set:

[0181] 3.1. Evaluation of the main efficacy indicators of Danshu Fuyuan Formula (granules) in the treatment of IMN: The study compared the changes in the levels of 24-hour urinary total protein (24hUTP) and albumin (ALB) in IMN subjects before and after treatment. The results showed that after 3 months and 6 months of treatment with Danshu Fuyuan granules, the level of 24hUTP in IMN subjects decreased significantly compared with that before treatment, and the difference was statistically significant (P < 0.01). After 6 months of treatment, the level of ALB increased slightly compared with that before treatment, and the difference was not statistically significant (P > 0.05). The specific results are shown in Table 4.

[0182] Table 4. Conditions of 24hUTP and ALB in 59 IMN subjects before and after treatment (x±s)

[0183]

[0184] Note: Compared with before treatment in the IMN group, *P < 0.05, **P < 0.01, the same below.

[0185] 3.2. Clinical remission rate of Danshu Fuyuan Formula (granules) in the treatment of IMN: After 3 months of treatment with Danshu Fuyuan granules, 3 out of 59 IMN subjects achieved complete remission, with a remission rate of 5.1%; 17 subjects achieved partial remission, with a remission rate of 28.8%, and the total remission rate was 33.9%. After 6 months of treatment, 5 subjects achieved complete remission, with a remission rate of 8.5%; 23 subjects achieved partial remission, with a remission rate of 39.0%, and the total remission rate was 47.5%.

[0186] 4. PPS analysis set:

[0187] 4.1. Evaluation of the main efficacy indicators of Danshu Fuyuan Formula (granules) in the treatment of IMN: The study compared the changes in the levels of 24hUTP and ALB in IMN subjects before and after treatment. The results showed that after 3 months of treatment with Danshu Fuyuan granules, the level of 24hUTP in IMN subjects decreased compared with that before treatment, and the difference was statistically significant (P < 0.05). The curative effect after 6 months of treatment was better than that after 3 months of treatment, and the downward trend of 24hUTP was more obvious, and the difference was statistically significant (P < 0.01). The level of ALB in 54 subjects increased slightly compared with that before treatment, and the difference was not statistically significant (P > 0.05). The specific results are shown in Table 5.

[0188] Table 5. Conditions of 24hUTP and ALB in 54 IMN subjects before and after treatment (x±s)

[0189]

[0190] 4.2 Evaluation of secondary efficacy indicators of Danshu Fuyuan Formula (granules) in the treatment of IMN: The study compared the changes in the levels of TP, BUN, CREA, and eGFR in IMN subjects before and after treatment. The results showed that there were no statistically significant differences in the levels of TP, BUN, CREA, and eGFR in IMN subjects after 3 months and 6 months of receiving Danshu Fuyuan Granules compared with those before treatment (P > 0.05). The specific results are shown in Table 6.

[0191] Table 6 Conditions of TP, BUN, CREA, and eGFR in 54 IMN subjects before and after treatment (x±s)

[0192]

[0193] 4.3 Clinical remission rate of Danshu Fuyuan Formula (granules) in the treatment of IMN: After 3 months of receiving Danshu Fuyuan Granules treatment, 3 IMN subjects achieved complete remission, with a remission rate of 5.6%; 17 subjects had partial remission, with a remission rate of 31.4%, and the total remission rate was 37.0%. After 6 months of treatment, 5 subjects achieved complete remission, with a remission rate of 9.3%; 24 subjects had partial remission, with a remission rate of 40.7%, and the total remission rate was 50.0%.

[0194] 4.4 TCM syndrome score and efficacy evaluation of Danshu Fuyuan Granules in the treatment of IMN: The study compared the changes in TCM syndrome scores in IMN subjects before and after treatment. The results showed that after 3 months and 6 months of receiving Danshu Fuyuan Granules treatment, the TCM syndrome scores of IMN subjects were significantly decreased (P < 0.01). The TCM syndrome scores after 6 months of treatment decreased more significantly than those after 3 months of treatment (P < 0.01). After 3 months of receiving Danshu Fuyuan Granules treatment, 2 IMN subjects achieved clinical cure, 0 had marked effect, 15 were effective, and 37 were ineffective, with a total effective rate of 31.5%; after 6 months of treatment, 6 subjects achieved clinical cure, 9 had marked effect, 29 were effective, and 10 were ineffective, with a total effective rate of 81.5%. The specific results are shown in Table 7.

[0195] Table 7 Results of TCM syndrome scores in 54 IMN subjects before and after treatment (x±s)

[0196]

[0197] Note: Compared with before treatment in the IMN group, *P < 0.05, **P < 0.01; compared between 6 months and 3 months of treatment, △ P < 0.05, △△ P < 0.01.

[0198] 4.5 One-year follow-up: Among the 54 IMN subjects, 25 subjects had 12-month follow-up information. The 24hUTP results of the 25 subjects were analyzed, and the specific results after 1 year are shown in Table 8.

[0199] Table 8 24hUTP results of 25 subjects (x±s)

[0200]

[0201] After 25 subjects received Danshu Fuyuan Granules for 3 months, 1 subject achieved complete remission, with a remission rate of 4%; 10 subjects had partial remission, with a remission rate of 40%, and the total remission rate was 44.0%. After 6 months of treatment, 3 subjects achieved complete remission, with a remission rate of 12%; 8 subjects had partial remission, with a remission rate of 32%, and the total remission rate was 44.0%. The results of the 1-year follow-up found that among the 25 subjects, 6 subjects achieved complete remission, with a remission rate of 24%; 12 subjects had partial remission, with a remission rate of 48%, and the total remission rate was 72%.

[0202] 4.6 Evaluation of the curative effect of Danshu Fuyuan Granules in the treatment of IMN by grouping

[0203] 4.6.1 Comparison of baseline data: The subjects were grouped according to whether they had used hormones or immunosuppressants in the past. The patients in the group who had used hormones or immunosuppressants in the past were those who had received standardized treatment in the Department of Nephrology of a tertiary hospital for more than 6 months and had not achieved clinical remission. The baseline levels between the two groups were compared.

[0204] According to whether they had used hormones or immunosuppressants in the past, 24 subjects had used hormones or immunosuppressants in the past, and 30 subjects had not. The gender, age, CKD stage, risk assessment, composition ratio of pathological stage, and laboratory test results (24hUTP, ALB, TP, BUN, CREA, eGFR) of the two groups of subjects were compared for differences, and there was no statistical significance (P>0.05). Therefore, the grouping comparison was comparable. The specific results are shown in Table 9.

[0205] Table 9 Comparison of baseline data of two groups of IMN subjects

[0206]

[0207]

[0208] 4.6.2、Previous use of hormones or immunosuppressants: Among the 24 subjects who had previously used hormones or immunosuppressants, 21 had used hormonal drugs, accounting for 87.5%; 8 had used cyclophosphamide, accounting for 33.3%; 8 had used cyclosporine, accounting for 33.3%; 6 had used tripterygium glycoside preparations, accounting for 25%; 4 had used tacrolimus, accounting for 16.7%; 2 had used leflunomide, accounting for 8.3%; 2 had used rituximab, accounting for 8.3%; and 1 had used mycophenolate mofetil, accounting for 4.2%.

[0209] 4.6.3、Comparison of clinical remission rates between the two groups: After 3 months of treatment with Danshu Fuyuan Granules, the clinical remission rate of the subjects who did not use hormones or immunosuppressants was 46.7%, and that of the subjects who had previously used hormones or immunosuppressants was 25%. There was no significant statistical difference in the clinical remission rates between the two groups (P > 0.05).

[0210] After 6 months of treatment with Danshu Fuyuan Granules, the clinical remission rate of the subjects who did not use hormones or immunosuppressants was 63.3%, and that of the subjects who had previously used hormones or immunosuppressants was 33.3%. There was a statistical difference in the clinical remission rates between the two groups (P < 0.05). As the treatment course extended, the efficacy of the subjects who did not use hormones or immunosuppressants was better. The specific results are shown in Table 10.

[0211] Table 10 Clinical remission rates of the two groups

[0212]

[0213] Note: Compared with those who had not used hormones or immunosuppressants, *P < 0.05 for those who had previously used hormones or immunosuppressants.

[0214] 5. Safety evaluation:

[0215] 5.1、Effect of Danshu Fuyuan Granules on safety indicators: Comparing the ALT and AST levels of IMN subjects at 3 months and 6 months of treatment, the results showed that there was no statistical difference in the ALT and AST results of the subjects before and after treatment (P > 0.05). The specific results are shown in Table 11.

[0216] Table 11 ALT and AST conditions of IMN subjects before and after treatment (x±s)

[0217]

[0218] 5.2、Complications during treatment: During the follow-up period, 4 cases had abnormal liver function and 1 case had respiratory tract infection. No serious adverse events occurred.

[0219] Application Example 3: Application of Danshu Fuyuan Formula in the Preparation of Drugs for Treating the Syndrome of Intermingled Phlegm and Blood Stasis in Vascular Dementia (VD) (using the granule obtained in Example 3 as the drug)

[0220] (I) Efficacy Verification of Danshu Fuyuan Formula (Bayesian Basket Trial)

[0221] 1. Research Subjects

[0222] Using the Bayesian basket trial research design, outpatients and inpatients who visited the Encephalopathy Department of Hubei Provincial Hospital of Traditional Chinese Medicine, the Hall of Traditional Chinese Medicine Masters, and the Department of Neurology of Wuhan First Hospital from January 2021 to December 2021 were selected. A total of 52 patients who met the diagnostic criteria and inclusion criteria for the syndrome of intermingled phlegm and blood stasis in VD were collected.

[0223] 2. Trial Scheme

[0224] On the basis of conventional Western medicine treatment, the subjects were given Danshu Fuyuan Formula granules (the formula granules were uniformly prepared and provided by China Resources Sanjiu Medical & Pharmaceutical Co., Ltd.), taken with warm boiled water, once in the morning and once in the evening, 1 bag each time.

[0225] If the patient had other underlying diseases, they continued to take the relevant symptomatic Western medicines (such as antihypertensive, lipid-lowering, hypoglycemic, antiplatelet aggregation drugs, etc.), but could not use other traditional Chinese medicines or Western medicines that had an impact on cognitive function.

[0226] The trial period of this study was 24 weeks. All enrolled patients collected the efficacy indicators before the intervention and at 12 weeks and 24 weeks (within ±7 days) after the intervention.

[0227] 3. Diagnostic Criteria

[0228] 3.1 Western Medicine Diagnostic Criteria

[0229] Diagnosis was made with reference to the diagnostic criteria for VD or vascular cognitive impairment published by Vas-Cog in 2014 and the "2018 Chinese Guidelines for the Diagnosis and Treatment of Dementia and Cognitive Impairment (I): Diagnostic Criteria for Dementia and Its Classification".

[0230] 1) Meeting the diagnostic criteria for dementia: For patients who were previously of normal intelligence but later developed cognitive decline (including impairment of memory, executive function, language, or visuospatial function) or abnormal mental behavior, which affects the patient's work or daily life and cannot be explained by delirium or other mental illnesses, a tentative diagnosis of dementia can be made. Cognitive decline or mental behavior impairment can be objectively confirmed by collecting the medical history and neuropsychological assessment, and at least two of the following five items must be met: a. A decline in memory and learning ability compared to before; b. Impairment of executive functions such as reasoning, judgment, and handling complex affairs; c. Impairment of visuospatial function; d. Impairment of language function (including listening, speaking, reading, and writing); e. Abnormal changes in the patient's behavior, personality, or demeanor. The above-mentioned functional impairments significantly affect the patient's daily living ability, occupation, or social interaction, and these impairments are not simply due to the physical disabilities caused by stroke resulting in a decline in daily living ability.

[0231] 2) Evidence of cerebrovascular lesions: Including relevant risk factors for vascular lesions, a history of stroke, focal neurological signs, and cerebrovascular lesions shown by imaging examinations. It is not necessary for all of the above evidence to be present simultaneously.

[0232] 3) There should be a causal relationship between dementia and cerebrovascular lesions: Clinically, by asking the patient's medical history, conducting physical examinations, and relevant laboratory and imaging examinations, it is determined whether there is a causal relationship between dementia and vascular factors. And the occurrence of dementia should be related to at least one cerebrovascular event in terms of time (with repeated similar cerebrovascular events, dementia shows a stepwise progression or fluctuates, and dementia still persists three months after the occurrence of the cerebrovascular event). Exclude other types of dementia (such as: Alzheimer's disease (AD), frontotemporal dementia (FTD), dementia with Lewy bodies (DLB), Parkinson's disease dementia (PDD), etc.).

[0233] 3.2、Traditional Chinese Medicine Diagnostic Criteria

[0234] 1) Main symptoms: Intellectual decline, abnormal behavior, slow reaction, dull expression, apathy, few words, mental confusion.

[0235] 2) Secondary symptom 1: Dizziness or heaviness, limb heaviness, chest fullness, obesity, coughing and expectorating phlegm.

[0236] 3) Secondary symptom 2: Headache like a stabbing pain, dull face and purple lips, purple and dull fingernails, hemiplegia of limbs, speech impairment.

[0237] 4) Tongue manifestation: Enlarged tongue with tooth marks, greasy tongue coating, purple and dull tongue with ecchymosis, and dilated and cyanotic sublingual veins.

[0238] 5) Pulse condition: Slippery or soft pulse, deep or unsmooth or taut pulse.

[0239] At least two of the above main symptoms (intellectual decline is essential), one item each from the secondary symptoms one and two, and one item each for the tongue and pulse manifestations are required for the diagnosis.

[0240] 4. Test Observation Indicators

[0241] 4.1 General Information

[0242] Demographic data: including name, age, gender, ethnicity, height, weight, marital status, education level, occupation, medical history, smoking and drinking history, etc.

[0243] Routine physical examination: such as body temperature, heart rate, respiration, pulse, blood pressure, etc.

[0244] Safety indicators: blood and urine routine, liver and kidney function, and electrocardiogram.

[0245] 4.2 Efficacy Indicators

[0246] Main indicators: The improvement of traditional Chinese medicine (TCM) clinical symptoms and signs in patients before and after treatment was evaluated using the TCM syndrome score scale (as shown in Table 12).

[0247] Table 12

[0248]

[0249]

[0250] Secondary indicators:

[0251] 1) Cognitive function: MMSE and MoCA scales were used to evaluate the changes in memory, orientation, calculation ability, etc. in patients before and after treatment.

[0252] 2) Activities of daily living (ADL): The ADL scale was used to evaluate the changes in the ADL of patients before and after treatment.

[0253] 5. Research Results

[0254] 5.1 General Information

[0255] From September 2020 to December 2021, a total of 52 subjects meeting the inclusion criteria were screened, and 2 cases were lost to follow-up, 1 case each for the first and second times.

[0256] 1) Distribution of patient gender, age, and disease duration

[0257] Among the 52 patients, 25 were male and 27 were female; the age of the patients was between 41 and 85 years old, with an average age of 67.83 ± 9.44 years; the disease duration of the patients was 6 to 37 months, with an average of 18.25 ± 9.62 months.

[0258] 2) Degree of the patient's illness, educational level, and distribution of co-existing diseases

[0259] Degree of the patient's illness (CDR classification): 35 cases with mild illness and 17 cases with moderate illness; Educational level of the patients: 4 illiterate, 15 with primary school education, and 33 with junior high school education or above; Conditions of co-existing diseases in the patients: 27 cases with hypertension, 8 cases with coronary heart disease, 13 cases with diabetes, 20 cases with hyperlipidemia, and 15 cases with hyperhomocysteinemia.

[0260] 5.2 Efficacy indicators

[0261] 1) Changes in the TCM syndrome scores of the patients

[0262] After treatment, the TCM syndrome scores of the patients were all lower compared with those before treatment, and the difference was statistically significant (P < 0.01), as shown in Table 13 specifically.

[0263] Table 13 Comparison of TCM syndrome scores (points) of patients before and after treatment in the Bayesian trial (x±s)

[0264]

[0265] Note: Comparison after treatment with before treatment: *P < 0.01.

[0266] 2) Changes in the MMSE scores of the patients

[0267] After treatment, the MMSE scores of the patients were all higher compared with those before treatment, and the difference was statistically significant (P < 0.01), as shown in Table 14 specifically.

[0268] Table 14 Comparison of MMSE scores (points) of patients before and after treatment in the Bayesian trial (x±s)

[0269]

[0270] Note: Comparison after treatment with before treatment: *P < 0.01.

[0271] 3) Changes in the MoCA scores of the patients

[0272] After treatment, the MoCA scores of the patients were all higher compared with those before treatment, and the difference was statistically significant (P < 0.01), as shown in Table 15 specifically.

[0273] Table 15 Comparison of MoCA scores (points) of patients before and after treatment in the Bayesian trial (x±s)

[0274]

[0275] Note: Comparison after treatment with before treatment: *P < 0.01.

[0276] 4) Changes in the ADL scores of the patients

[0277] After treatment, the ADL scores of the patients were all lower than those before treatment, and the difference was statistically significant (P < 0.01), as shown in Table 16 specifically.

[0278] Table 16 Comparison of ADL scores (points) of patients in the Bayesian trial before and after treatment (x±s)

[0279]

[0280] Note: Comparison after treatment with before treatment: *P<0.01.

[0281] 5) Safety indicators

[0282] During the whole trial process, 2 cases were lost to follow-up, 1 case each in March and June, all due to loss to follow-up.

[0283] All patients in this study underwent blood, urine routine, liver and kidney function, and electrocardiogram examinations before and after treatment. No abnormal changes in the above indicators were found in both groups of patients. Among the 50 patients who completed the trial, 2 patients (incidence rate 4%) complained of digestive tract reactions (nausea, discomfort in the epigastric region) after taking the medicine. The patients were instructed to take the medicine more than half an hour after meals, and no special drug treatment was given. The symptoms of the patients disappeared spontaneously.

[0284] 6. Discussion

[0285] The inventors have long-term research and found that the location of vascular dementia is in the brain, which is closely related to the kidneys, heart, spleen, and liver, and is also related to qi, blood, and body fluids. Qi and blood are the material basis for the mental activities of the brain. Therefore, the deficiency of primordial qi and essence and blood, resulting in the reduction of marrow and atrophy of the brain, is the root cause of this disease; phlegm turbidity and stasis blood are not only the pathological products generated by the deficiency of various zang-organs, but also the pathogenic factors leading to the dysfunction of the mental mechanism. The two often exist simultaneously, forming the syndrome of phlegm-stasis mutual binding. Clinically, the method of treating phlegm and stasis simultaneously is often used to treat this disease. The present invention believes that "phlegm-stasis mutual binding" is the key pathogenesis of cognitive function impairment in patients with vascular dementia. Therefore, taking "phlegm-stasis mutual binding" as the entry point, the treatment principle of "tonifying qi, resolving phlegm, promoting blood circulation, clearing heat, and dissipating nodules" (i.e., Danshu Fuyuan Prescription) is proposed, and syndrome differentiation and treatment are carried out on the patients, and the significant curative effect is proved by the above results.

[0286] 7. Conclusion

[0287] The results of this study showed that the basic formula of "tonifying qi, resolving phlegm, activating blood circulation, clearing heat and dissipating stasis" (Danshu Fuyuan Formula) could effectively reduce the TCM syndrome scores of VD patients and improve the TCM syndromes of the patients (the average score before treatment was 30.27, 19.00 after 3 months of treatment, and 14.96 after 6 months of treatment); from the comparison of the MMSE scores of the patients before and after treatment, it was found that it had a therapeutic effect on improving the cognitive function of the patients (the average score before treatment was 17.24, 21.02 after 3 months of treatment, and 22.86 after 6 months of treatment); from the comparison of the MoCA scores of the patients before and after treatment, it was found that it had a therapeutic effect on improving the cognitive function of the patients (the average score before treatment was 16.58, 19.53 after 3 months of treatment, and 21.48 after 6 months of treatment); from the comparison of the ADL scores of the patients before and after treatment, it was found that it had a therapeutic effect on improving the daily living ability of the patients (the average score before treatment was 38.96, 32.31 after 3 months of treatment, and 28.30 after 6 months of treatment); the incidence of adverse reactions (gastrointestinal reactions, without special treatment and disappeared spontaneously) was 4%, indicating that Danshu Fuyuan Formula had good safety.

[0288] As described above, it is only the specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present invention can easily think of changes or substitutions, which should be covered within the protection scope of the present invention. Therefore, the protection scope of the present invention should be subject to the protection scope of the claimed rights.

Claims

1. A Danshu Fuyuan formula for treating the syndrome of intermingled phlegm and stasis, characterized in that, The raw materials of the Danshu Fuyuan Prescription are in a weight ratio of: 25-35 parts of Salvia miltiorrhiza, 10-15 parts of Atractylodes macrocephala, 25-35 parts of Astragalus membranaceus, 10-20 parts of Lonicera japonica, 10-20 parts of Polygonum cuspidatum, 10-20 parts of Curcuma aromatica, 10-15 parts of Poria cocos, 25-35 parts of Coix lacryma-jobi and 10-15 parts of Notopterygium incisum; The phlegm-stasis mutual binding syndrome is one of the phlegm-stasis mutual binding syndromes of coronary heart disease, idiopathic membranous nephropathy, and vascular dementia.

2. The Danshu Fuyuan formula according to claim 1, characterized in that, The raw materials of the Danshu Fuyuan Prescription are in a weight ratio of: 30 parts of Salvia miltiorrhiza, 12 parts of Atractylodes macrocephala, 30 parts of Astragalus membranaceus, 15 parts of Lonicera japonica, 15 parts of Polygonum cuspidatum, 15 parts of Curcuma aromatica, 12 parts of Poria cocos, 30 parts of Coix lacryma-jobi and 12 parts of Notopterygium incisum.

3. A preparation method of the Danshu Fuyuan formula according to claim 1 or 2, characterized in that, The preparation method is as follows: Crush the raw materials of the Danshu Fuyuan Prescription and then mix them to obtain a composition; Or, mix the raw materials of the Danshu Fuyuan Prescription and then crush them to obtain a composition; Or, extract and refine the raw materials of the Danshu Fuyuan Prescription according to the conventional process to obtain one of tablets, capsules, granules, pills, powders or oral liquids.

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