Traditional Chinese medicine composition and use thereof

Abstract

The present disclosure relates to the field of drugs and health foods. Disclosed is a traditional Chinese medicine composition comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil. Experiments show that the traditional Chinese medicine composition has a scientific formulation, synergistic functions, without toxic and side effects, and not only has nutritional value but also inhibits the growth of tumor. Comparing with individual components, the composition significantly increases the percentages of CD4.sup.+ and CD8.sup.+ cells in lymphocytes of the tumor stroma, enhances body immune function, restores normal immune surveillance function, decreases VEGF and TGF-β1 positive cells in tumor tissue, assists in treating tumor, increases antitumor effects of chemotherapy drugs, improves immune status of body, has the function of restoring normal immune surveillance, preventing and assisting in treating tumor.

Claims

1. A method of inhibiting the growth of tumor, increasing the percentage of CD.sub.4.sup.+ and CD.sub.8.sup.+ cells in lymphocytes of tumor stroma, decreasing VEGF and TGF-β1 positive cells in tumor tissue, enhancing immunity, preventing and/or assisting in treating tumor, and/or restoring normal immune function, comprising administrating an effective amount of a traditional Chinese medicine composition to a subject in need thereof; wherein the traditional Chinese medicine composition comprises GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil, wherein the weight ratio of GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil is (1 to 5):(1 to 5):(1 to 5):(1 to 5):(1 to 5).

2. The method according to claim 1, wherein the weight ratio of GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil is (1 to 2):(1 to 2):(1 to 2):(1 to 2):(1 to 2).

3. The method according to claim 1, wherein the weight ratio of GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil is 1:1:1.5:1.5:1.5.

4. The method according to claim 1, where in the traditional Chinese medicine composition is in the form of a health food or a medicine.

5. The method according to claim 4, wherein the health food or the medicine further comprises a pharmaceutically acceptable excipient or a food acceptable excipient.

Description

DETAILED DESCRIPTION

(1) The present disclosure provides a traditional Chinese medicine composition and the application thereof. One of ordinary skill in the art can learn from the contents herein and improve the process parameters appropriately. In particular, it shall be noted that all the similar substitutions and modifications are apparent to one of ordinary skill in the art and are to be considered within the scope of the present invention. The method and product of the present invention have been described with preferred examples. It is apparent that one of the ordinary skill in the art can make change or modify the combination to the method and product of the present invention without departing from the spirit, scope and spirit of the invention, therefore realizing and applying the techniques of the present invention.

(2) In order to understand the present disclosure further, the technical solutions in the embodiments of the present disclosure will be described clearly and completely herein in conjunction with Examples of the present disclosure. Apparently, the described examples are only a part of Examples of the present disclosure, rather than all examples. Based on Examples in the present disclosure, all of other examples, made by one of ordinary skill in the art without any creative efforts, fall into the protection scope of the present disclosure.

(3) Without special illustration, all the reagents in Examples of the present disclosure are commercial products, which can be purchased on the market.

Example 1 Preparation of GANODERMA Polysaccharide

(4) GANODERMA polysaccharide of the present disclosure was prepared by the following method. GANODERMA was pulverized, and water was added at a weight ratio of GANODERMA:water=1:10. Extraction was performed for 2 hours under conditions of heating and stirring at 70° C. The filter residue was collected and continuously subjected to extraction for twice according to the solid-to-liquid ratio, temperature and duration above. Extracts of the three extractions were combined, concentrated under reduced pressure and centrifuged to obtain a clarified concentrate. 95% alcohol which was 7 times the volume of the concentrate was added to the clarified concentrate, mixed, and the mixture was held for precipitation over night at 4° C. The mixture was centrifuged to collect the precipitates, and the precipitates were subjected to vacuum freeze-drying to obtain the GANODERMA polysaccharide.

Example 2 Preparation of MORINDAE OFFICINALIS RADIX Polysaccharide

(5) MORINDAE OFFICINALIS RADIX polysaccharide of the present disclosure was prepared by the following method. MORINDAE OFFICINALIS RADIX was pulverized, and water was added at a weight ratio of MORINDAE OFFICINALIS RADIX:water=1:5. Extraction was performed for 3 hours under conditions of heating and stirring at 75° C. The filter residue was collected and continuously subjected to extraction for twice according to the solid-to-liquid ratio, temperature and duration above. Extracts of the three extractions were combined, concentrated under reduced pressure and centrifuged to obtain a clarified concentrate. 95% alcohol which was 9 times the volume of the concentrate was added to the clarified concentrate, mixed, and the mixture was held for precipitation over night at 4° C. The mixture was centrifuged to collect the precipitates, and the precipitates were subjected to vacuum freeze-drying to obtain the MORINDAE OFFICINALIS RADIX polysaccharide.

Example 3 Preparation of POLYGONATI RHIZOMA Polysaccharide

(6) POLYGONATI RHIZOMA polysaccharide of the present disclosure was prepared by the following method. POLYGONATI RHIZOMA was pulverized, and water was added at a weight ratio of POLYGONATI RHIZOMA:water=1:5. Extraction was performed for 3 hours under conditions of heating and stirring at 75° C. The filter residue was collected and continuously subjected to extraction for twice according to the solid-to-liquid ratio, temperature and duration above. Extracts of the three extractions were combined, concentrated under reduced pressure and centrifuged to obtain a clarified concentrate. 95% alcohol which was 9 times the volume of the concentrate was added to the clarified concentrate, mixed, and the mixture was held for precipitation over night at 4° C. The mixture was centrifuged to collect the precipitates, and the precipitates were subjected to vacuum freeze-drying to obtain the POLYGONATI RHIZOMA polysaccharide.

Example 4 Preparation of ANGELICAE SINENSIS RADIX Oil

(7) ANGELICAE SINENSIS RADIX oil of the present disclosure was prepared by the following method. 400 g of ANGELICAE SINENSIS RADIX fine powder (10 meshes) was weighted, and water which was 15 times the weight of the fine powder was added and mixed to reach a solid-to-liquid ratio of 1:15. The fine powder was soaked for 5 hours and then subjected to steam distillation extraction. Sodium chloride (80 g) which was ⅕ time the weight of ANGELICAE SINENSIS RADIX was added during the extraction. The extraction temperature was from 140 to 150° C., the vapor flux was 2 L/h, and the duration was 8 hours. The ANGELICAE SINENSIS RADIX oil obtained has a light yellow color, with a special aroma of ANGELICAE SINENSIS RADIX and a bitter taste, and a yield from 0.2 to 0.3%, which was preserved in a refrigerator at −20° C.

Example 5 Preparation of MYRISTICAE SEMEN Oil

(8) MYRISTICAE SEMEN oil of the present disclosure was prepared by the following method. 400 g of MYRISTICAE SEMEN fine powder (20 meshes) was weighted, and water which was 15 times the weight of the fine powder was added and mixed to reach a solid-to-liquid ratio of 1:15. The fine powder was soaked for 1 hour and then subjected to steam distillation extraction. The extraction temperature was from 130 to 140° C., the vapor flux was 2 L/h, and the duration was 6 hours. The MYRISTICAE SEMEN oil obtained has a yellow color, with a spicy and intense flavor and a taste of musk, and a yield from 10 to 12%, which was preserved in a refrigerator at −20° C.

Example 6 Preparation of the Traditional Chinese Medicine Composition

(9) GANODERMA polysaccharide prepared in Example 1, MORINDAE OFFICINALIS RADIX polysaccharide prepared in Example 2, POLYGONATI RHIZOMA polysaccharide prepared in Example 3, ANGELICAE SINENSIS RADIX oil prepared in Example 4 and MYRISTICAE SEMEN oil prepared in Example 5 were mixed at a weight ratio of 1:1:1.5:1.5:1.5 to obtain a traditional Chinese medicine composition comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil.

Example 7 Preparation of the Traditional Chinese Medicine Composition

(10) GANODERMA polysaccharide prepared in Example 1, MORINDAE OFFICINALIS RADIX polysaccharide prepared in Example 2, POLYGONATI RHIZOMA polysaccharide prepared in Example 3, ANGELICAE SINENSIS RADIX oil prepared in Example 4 and MYRISTICAE SEMEN oil prepared in Example 5 were mixed at a weight ratio of 1:5:4:2:4 to obtain a traditional Chinese medicine composition comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil.

Example 8 Preparation of the Traditional Chinese Medicine Composition

(11) GANODERMA polysaccharide prepared in Example 1, MORINDAE OFFICINALIS RADIX polysaccharide prepared in Example 2, POLYGONATI RHIZOMA polysaccharide prepared in Example 3, ANGELICAE SINENSIS RADIX oil prepared in Example 4 and MYRISTICAE SEMEN oil prepared in Example 5 were mixed at a weight ratio of 5:1:1:5:1 to obtain a traditional Chinese medicine composition comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil.

Example 9 Preparation of the Traditional Chinese Medicine Composition

(12) GANODERMA polysaccharide prepared in Example 1, MORINDAE OFFICINALIS RADIX polysaccharide prepared in Example 2, POLYGONATI RHIZOMA polysaccharide prepared in Example 3, ANGELICAE SINENSIS RADIX oil prepared in Example 4 and MYRISTICAE SEMEN oil prepared in Example 5 were mixed at a weight ratio of 1:2:1:2:1 to obtain a traditional Chinese medicine composition comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil.

Example 10 Preparation of the Traditional Chinese Medicine Composition

(13) GANODERMA polysaccharide prepared in Example 1, MORINDAE OFFICINALIS RADIX polysaccharide prepared in Example 2, POLYGONATI RHIZOMA polysaccharide prepared in Example 3, ANGELICAE SINENSIS RADIX oil prepared in Example 4 and MYRISTICAE SEMEN oil prepared in Example 5 were mixed at a weight ratio of 2:4:5:1:5 to obtain a traditional Chinese medicine composition comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil.

Example 11 Test the Function of the Traditional Chinese Medicine Composition on Preventing or Assisting in Treating Tumor

(14) GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil, MYRISTICAE SEMEN oil and the traditional Chinese medicine compositions obtained in examples 1 to 10 above were subjected to cell experiments by the inventors. The experiments were shown hereinafter.

(15) 1. Experiment Materials

(16) 1.1 Experiment Equipment and Reagents

(17) Flow cytometer: BD, Calibo.

(18) Analytical balance: Beijing Sartorius Co., Ltd., Sartorius BS124S type.

(19) Electronic balance: Beijing Sartorius Co., Ltd., Sartorius BL1500 type.

(20) Carbon dioxide constant-temperature cell incubator (Nuaire).

(21) Low-temperature high-speed centrifuge (Hunan Xingke TGL-16G).

(22) The CD4.sup.+, CD8.sup.+ and antibody for flow cytometry were purchased from BD.

(23) Lymphocyte separation solution (Guanzhou Zhanchen Biological Technology Co., Ltd., China)

(24) Fluorescein isothiocyanate labeled anti-mouse CD4 and CD8a monoclonal antibody (BioLegend, Inc., U.S.A.)

(25) VEGF and TGF-β1 immunocytochemistry kit (Beijing Zhongshan Biological Technology Co., Ltd., China)

(26) 1.2 Experimental Drugs

(27) The GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil and the traditional Chinese medicine compositions prepared in examples 1 to 10, store at 4° C. and avoid light.

(28) 1.3 Experimental Animals

(29) Female SPF mice were purchased from Laboratory Animal Research Center, Institute of Biophysics, Chinese Academy of Sciences. The mice were housed under conditions of 4 mice/box, temperature 20 to 25° C., RH 40 to 70% and 12 h:12 h day-night intermittently illumination. The mice have free access to food and drinking water, and the drinking water was distilled water prepared by the Laboratory Animal Research Center. The mice were divided into normal group, model group and groups of examples 1 to 10, 8 mice per group.

(30) 2. Experimental Method

(31) 2.1 Test of Inhibition Rate of Tumor

(32) Colon cancer cell line CT26 was subcutaneously inoculated to Balb/c mice at an inoculation dosage of 1×10.sup.6/mouse. 12 days later, the mice were administered by intragastric gavage with the compositions of examples 1 to 10. The administration was performed once a day at a dosage of 500 mg/kg/day. The normal group and the model group were administered by intragastric gavage with distilled water. On the 33.sup.th day, 24 hours after the last administration, blood samples were collected from the hearts and the mice were sacrificed by cervical dislocation. The entire tumors were peeled off and weighed, and the inhibition rate of tumor was calculated.
The inhibition rate=(mean tumor weight of the model group−mean tumor weight of the administration group)/mean tumor weight of the model group×100%.
2.2 Test of CD4.sup.+ and CD8.sup.+ in Lymphocytes of Tumor Stroma and Intervention of the Traditional Chinese Medicine Composition

(33) Tumors were taken from the mice by surgery under asepsis condition. The tumors were weighted by an analytical balance and the tumors with a weight over 2 g were selected. A 1 mm.sup.3 block was constructed from the selected tumor tissues, disposed in a RPMI-1640 culture medium containing 0.05% of collagenase, 0.02% of DNase, 0.01% of hyaluronidase and 10% of calf serum, and subjected to digestion under magnetic stirring at 4° C. overnight. The tissues were screened with a 200 meshes steel sieve. All the single cells were collected, subjected to low speed centrifugation (1500 r/min) for 15 minutes, and washed with Hanks solution for twice. The cell concentration was adjusted to about 1×10.sup.6 cells/mL, i.e., tumor infiltrating lymphocyte.

(34) 2 EP tubes were prepared for each test sample. 50 μL cell suspension was added to each tube and the total volume was adjusted to 100 μL with a PBS buffer to reach a final concentration of 5×10.sup.5 cells/mL. In one tube, 0.25 μg of anti-mouse CD4 monoclonal antibody and 0.2 μg of anti-mouse CD8a monoclonal antibody labeled with different fluorescein were added, and the other tube was set as the blank control tube. The tubes were incubated at room temperature for 30 minutes by avoiding light. The cells were washed with PBS and tested by a flow cytometer.

(35) 2.3 Test of expression rate of VEGF and TGF-β1 positive cells in tumor tissue Immunohistochemistry methods were used according to the protocols. The results were determined by the appearance of brown color in cytoplasm of the VEGF and TGF-β1 staining positive cells.
2.4 Statistical Method

(36) All the experimental data in the experiments was statistically analyzed with software SPSS19.0. All the measurement data was expressed in the form of mean±standard deviation (−x±s), and t Test was performed. And the enumeration data was tested with χ2 Test. When P<0.05, the difference is statistically significant.

(37) 3. Experimental Results

(38) 3.1 Tumor Inhibition Rate in Mice

(39) TABLE-US-00001 TABLE 1 Inhibition effect of the samples on tumor growth in mice (n = 8) Group Normal Model Group Group Example 1 Example 2 Example 3 Example 4 Tumor — — 31.36 35.27 32.39 33.20 Inhibition Rate (%) Group Example Example 5 Example 6 Example 7 Example 8 Example 9 10 Tumor 32.65 45.37 40.16 39.82 42.81 41.12 Inhibition Rate (%)

(40) As shown in Table 1, all the groups of examples 1 to 10 have inhibition effects on the growth of mice tumor. Therein, inhibition rate of Example 6 was the highest, 45.37%; the second was Example 9, 42.81%; while Example 1 has the least inhibition rate, 31.36%. The results indicated that both the individual components and the traditional Chinese medicine compositions were capable to inhibit the growth of tumor and played a role in preventing and assisting in treating tumor. However, the effects of the traditional Chinese medicine compositions were better than that of the individual components.

(41) 3.2 Effects on CD4.sup.+, CD8.sup.+ Cells in Lymphocyte of Tumor Stroma.

(42) TABLE-US-00002 TABLE 2 Effects of examples on CD4.sup.+, CD8.sup.+ cells in mice (n = 8) Group CD4.sup.+(%) CD8.sup.+(%) Normal Group 45.82 ± 3.09 27.50 ± 1.21.sup.  Model Group  13.01 ± 1.13*  .sup. 7.35 ± 0.69* Example 1 16.94 ± 1.79 .sup. 8.35 ± 0.57 Example 2 .sup. 18.12 ± 2.47.sup.# .sup. 9.19 ± 0.81 Example 3 17.24 ± 5.02 .sup. 8.99 ± 0.92 Example 4 17.46 ± 2.92  9.25 ± 0.47.sup.# Example 5 17.46 ± 2.37 .sup. 8.28 ± 1.14 Example 6 .sup. 26.46 ± 2.37.sup.# 14.28 ± 1.14.sup.# Example 7 .sup. 24.67 ± 2.11.sup.# 13.09 ± 1.16.sup.# Example 8 24.14 ± 6.22 12.78 ± 2.07.sup.# Example 9 .sup. 25.09 ± 3.17.sup.# 13.99 ± 1.45.sup.# Example 10 24.75 ± 5.32 12.95 ± 2.11.sup.# Comment: comparing with the normal group, *P < 0.05; and comparing with the model group, .sup.#P < 0.05.

(43) As shown in Table 2, each of the groups of examples 1 to 10 increased the percentages of CD4.sup.+ and CD8.sup.+ cells in lymphocyte of tumor stroma in some degree. In addition, the CD4.sup.+ and CD8.sup.+ differences between examples 6 to 10 and the model group were statistically significant (P<0.05). Therein, detection values of both CD4.sup.+ and CD8.sup.+ cells in the group of Example 6 were the highest, (26.46±2.37)% and (14.28±1.14)%, respectively. The second was Example 9, with detection values of (25.09±3.17)% and (13.99±1.45)%, respectively. The results indicated that both the individual components and the traditional Chinese medicine compositions increased the percentages of CD4.sup.+ and CD8.sup.+ cells and improved the immune status of body, which could be used to produce health products and foods for preventing and assisting in treating tumor. However, the effects of the traditional Chinese medicine compositions were better than that of the individual components.

(44) 3.3 Effects on Expression Rate of VEGF and TGF-β1 Positive Cells in Tumor

(45) TABLE-US-00003 TABLE 3 Effects of samples on VEGF and TGF-β1 positive cells in tumor Group VEGF (%) TGF-β 1 (%) Normal Group 31.64 ± 3.09 35.76 ± 1.21 Model Group  75.43 ± 9.21*  70.36 ± 8.25* Example 1 69.28 ± 9.01 67.48 ± 7.92 Example 2 63.79 ± 2.47 65.02 ± 0.81 Example 3 65.47 ± 5.02 66.41 ± 0.92 Example 4  66.1 ± 2.92 66.73 ± 0.47 Example 5 65.31 ± 5.11 66.17 ± 0.83 Example 6  .sup. 58.49 ± 5.87 .sup.#  .sup. 60.12 ± 3.28 .sup.# Example 7  .sup. 60.99 ± 2.11 .sup.#  .sup. 61.12 ± 1.16 .sup.# Example 8 61.62 ± 6.22  .sup. 61.63 ± 2.07 .sup.# Example 9  .sup. 59.11 ± 3.17 .sup.#  .sup. 60.78 ± 1.45 .sup.# Example 10 61.15 ± 5.32  .sup. 61.15 ± 2.11 .sup.# Comment: comparing with the normal group, *P < 0.05; and comparing with the model group, .sup.#P < 0.05.

(46) As shown in Table 3, each of the groups of examples 1 to 10 decreased the expression rate of VEGF and TGF-β1 positive cells in mice tumor. Therein, the differences of VEGF and TGF-β1 positive cells between examples 6-10 and the model groups were statistically significant (P<0.05), while the differences between examples 1 to 5 and the model group were not significant. Detection values of both the VEGF and TGF-β1 positive cells of the group of Example 6 were the least, (58.49±5.87)% and (60.12±3.28)%, respectively. The second least was Example 9, (59.11±3.17)% and (60.78±1.45)%, respectively. The results indicated that both the individual components and the traditional Chinese medicine compositions decreased the expression rate of VEGF and TGF-β1 positive cells in mice tumor, inhibited the expression of immune inhibitive factors in local tumor tissue, therefore delaying the development of tumor and improving the immune status of body. They could be used in preparing health products and foods, for preventing and assisting in treating tumor. However, the effects of the traditional Chinese medicine compositions were better than that of the individual components.

(47) It can be concluded from the above results that the traditional Chinese medicine compositions comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil can inhibit the growth of tumors, improve percentages of CD4.sup.+ and CD8.sup.+ cells in the lymphocytes of the tumor stroma, and decrease the expression rate of VEGF and TGF-β1 positive cells in mice tumor. The role was to inhibit the growth of tumor cells, recover normal immune surveillance and prevent immune escape of the tumor cells. Therein, the traditional Chinese medicine composition which was prepared in Example 6 and consisted of GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil has the best effects on assisting in treating tumor and restoring the normal immune surveillance of body, and the effects were better than that of the individual components (P<0.05).

Example 12 Compositions Comprising GANODERMA Polysaccharide, MORINDAE OFFICINALIS RADIX Polysaccharide, POLYGONATI RHIZOMA Polysaccharide, ANGELICAE SINENSIS RADIX Oil and MYRISTICAE SEMEN Oil with Different Ratios, and Effects Thereof

(48) GANODERMA polysaccharide prepared in Example 1, MORINDAE OFFICINALIS RADIX polysaccharide prepared in Example 2, POLYGONATI RHIZOMA polysaccharide prepared in Example 3, ANGELICAE SINENSIS RADIX oil prepared in Example 4 and MYRISTICAE SEMEN oil prepared in Example 5 were mixed according to the different weight ratios shown in the Table 4. The compositions were subjected to the test methods of Example 11 for the effects on preventing and/or assisting in treating tumors. The scores of efficacy for restoring normal immune surveillance of the body were obtained (100 point scale, the efficacy for restoring normal immune surveillance by the traditional Chinese medicine composition of Example 6 with a weight ratio of 1:1:1.5:1.5:1.5 was recorded as 100 points, and the percentages of other mixing ratios compared to the function test result of Example 6 are recorded as the corresponding scores).

(49) TABLE-US-00004 TABLE 4 Compositions comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil with different weight ratios, and effects thereof Ratio of Components 1:5:5:1:5 1:5:4:2:4 4:2:5:1:1 2:1:1:2:1 1:2:2:1:1.5 1:1.5:1:1.5:1 1.5:1:1.5:1:1.5 Score of Restoring 65 70 74 80 83 91 93 Normal Immune Surveillance

(50) The results showed that comparing with using the individual components alone, the compositions comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil with a mixing ratio of (1˜5):(1˜5):(1˜5):(1˜5):(1˜5) showed more significant effects on restoring normal immune surveillance of body. When the mixing ratio was (1˜2):(1˜2):(1˜2):(1˜2):(1˜2), the effects were more significant.

Example 13 A Polysaccharide Composition Comprising GANODERMA Polysaccharide, MORINDAE OFFICINALIS RADIX Polysaccharide and POLYGONATI RHIZOMA Polysaccharide with a Ratio of 1:1:1.5, and Effects Thereof

(51) GANODERMA polysaccharide prepared in Example 1, MORINDAE OFFICINALIS RADIX polysaccharide prepared in Example 2 and POLYGONATI RHIZOMA polysaccharide prepared in Example 3 were mixed in the best weight ratio of 1:1:1.5. The composition was subjected to the test methods of Example 11 for the effects on preventing and/or assisting in treating tumors. The scores of efficacy for restoring normal immune surveillance of the body were obtained (100 point scale, the efficacy for restoring normal immune surveillance by the traditional Chinese medicine composition of Example 6 with a weight ratio of 1:1:1.5:1.5:1.5 was recorded as 100 points, and the percentages of other mixing ratios compared to the function test result of Example 6 are recorded as the corresponding scores).

(52) TABLE-US-00005 TABLE 5 Compositions comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil with different ratios, and effects thereof Ratio of Components 1:1:1.5:0:0 1:5:5:1:5 1:5:4:2:4 4:2:5:1:1 2:1:12:1 1:2:2:1:1.5 1:1.5:1:1.5:1 1.5:1:1.5:1:1.5 Score of 47 65 70 74 80 83 91 93 Restoring Normal Immune Surveillance

(53) The results showed that comparing with using the polysaccharide composition alone, the compositions comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil with a mixing ratio of (1˜5):(1˜5):(1˜5):(1˜5):(1˜5) showed more significant effects on restoring normal immune surveillance of body.

Example 14 A Composition Comprising ANGELICAE SINENSIS RADIX Oil and MYRISTICAE SEMEN Oil with a Ratio of 1:1, and Effects Thereof

(54) ANGELICAE SINENSIS RADIX oil prepared in Example 4 and MYRISTICAE SEMEN oil prepared in Example 5 were mixed in the best weight ratio of 1:1. The composition was subjected to the test methods of Example 11 for the effects on preventing and/or assisting in treating tumors. The scores of efficacy for restoring normal immune surveillance of the body were obtained (100 point scale, the efficacy for restoring normal immune surveillance by the traditional Chinese medicine composition of Example 6 with a weight ratio of 1:1:1.5:1.5:1.5 was recorded as 100 points, and the percentages of other mixing ratios compared to the function test result of Example 6 are recorded as the corresponding scores).

(55) TABLE-US-00006 TABLE 6 Compositions comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil with different ratios, and effects thereof Ratio of Components 0:0:0:1:1 1:5:5:1:5 1:5:4:2:4 4:2:5:1:1 2:1:1:2:1 1:2:2:1:1.5 1:1.5:1:1.5:1 1.5:1:1.5:1:1.5 Score of Restoring 41 65 70 74 80 83 91 93 Normal Immune Surveillance

(56) The results showed that comparing with using the oil composition alone, the compositions comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil with a mixing ratio of (1˜5):(1˜5):(1˜5):(1˜5):(1˜5) showed more significant effects on restoring normal immune surveillance of body.

Example 15 Compositions Comprising GANODERMA Polysaccharide, MORINDAE OFFICINALIS RADIX Polysaccharide, POLYGONATI RHIZOMA Polysaccharide, ANGELICAE SINENSIS RADIX Oil and MYRISTICAE SEMEN Oil with Different Ratios, and Effects Thereof

(57) GANODERMA polysaccharide prepared in Example 1, MORINDAE OFFICINALIS RADIX polysaccharide prepared in Example 2, POLYGONATI RHIZOMA polysaccharide prepared in Example 3, ANGELICAE SINENSIS RADIX oil prepared in Example 4 and MYRISTICAE SEMEN oil prepared in Example 5 were mixed according to the weight ratios shown in Table 7 (each composition lacked one or two components). The compositions were subjected to the test methods of Example 11 for the effects on preventing and/or assisting in treating tumors. The scores of efficacy for restoring normal immune surveillance of the body were obtained (100 point scale, the efficacy for restoring normal immune surveillance by the traditional Chinese medicine composition of Example 6 with a weight ratio of 1:1:1.5:1.5:1.5 was recorded as 100 points, and the percentages of other mixing ratios compared to the function test result of Example 6 are recorded as the corresponding scores).

(58) TABLE-US-00007 TABLE 7 Compositions comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil with different ratios, and effects thereof Ratio of Components 1:5:5:1:5 1:5:4:2:4 4:2:5:1:1 2:1:1:2:1 1:2:2:1:1.5 1:1.5:1:1.5:1 1.5:1:1.5:1:1.5 Score of 65 70 74 80 83 91 93 Restoring Normal Immune Surveillance Ratio of Components 1:1:1.5:0:1.5 1:0:0:1.5:1.5 0:0:1:1:0 1:1:1.5:1.5:0 0:0:1:1:1 0:1:0:1.5:1.5 0:1:0:0:1.5 Score of 61 60 56 58 53 60 54 Restoring Normal Immune Surveillance

(59) The results showed that comparing with the compositions that lacked one or two components, the compositions comprising GANODERMA polysaccharide, MORINDAE OFFICINALIS RADIX polysaccharide, POLYGONATI RHIZOMA polysaccharide, ANGELICAE SINENSIS RADIX oil and MYRISTICAE SEMEN oil with a mixing ratio of (1˜5):(1˜5):(1˜5):(1˜5):(1˜5) showed more significant effects on restoring normal immune surveillance of body. When the mixing ratio was (1˜2):(1˜2):(1˜2):(1˜2):(1˜2), the effects were more significant.