TRADITIONAL CHINESE MEDICINE COMPOSITION FOR TREATING DISEASES CAUSED BY HUMAN PAPILLOMAVIRUS, PREPARATION METHOD AND USE THEREOF

Abstract

The present invention discloses a traditional Chinese medicine composition for treating diseases caused by human papillomavirus, a preparation method and use thereof. The traditional Chinese medicine composition comprises: 15-60 parts of Rhizoma Smilacis Glabrae, 7.5-30 parts of Sophora Flavescens, 10-40 parts of Herba Hedyotidis Diffusae, 10-40 parts of Herba Taraxaci, 6-24 parts of Dysosma Versipellis, 7.5-30 parts of Nidus Vespae, 10-40 parts of Cortex Phellodendri, 2.5-10 parts of Fructus Bruceae, 7.5-30 parts of Fructus Cnidii, 3-12 parts of Radix Glycyrrhizae, 7.5-30 parts of Flos Lonicerae, 7.5-30 parts of Folium Isatidis, 5-20 parts of Radix Semiaquilegiae, and 1000-2000 parts of 75% ethanol. The traditional Chinese medicine composition has good curative effect and good safety.

Claims

1. A traditional Chinese medicine composition, comprising the following pharmaceutical ingredients in parts by weight: 15-60 parts of Rhizoma Smilacis Glabrae, 7.5-30 parts of Sophora Flavescens, 10-40 parts of Herba Hedyotidis Diffusae, 10-40 parts of Herba Taraxaci, 6-24 parts of Dysosma Versipellis, 7.5-30 parts of Nidus Vespae, 10-40 parts of Cortex Phellodendri, 2.5-10 parts of Fructus Bruceae, 7.5-30 parts of Fructus Cnidii, 3-12 parts of Radix Glycyrrhizae, 7.5-30 parts of Flos Lonicerae, 7.5-30 parts of Folium Isatidis, 5-20 parts of Radix Semiaquilegiae, and 1000-2000 parts of 75% ethanol.

2. The traditional Chinese medicine composition according to claim 1, wherein the composition comprises the following pharmaceutical ingredients in parts by weight: 38 parts of Rhizoma Smilacis Glabrae, 19 parts of Sophora Flavescens, 30 parts of Herba Hedyotidis Diffusae, 30 parts of Herba Taraxaci, 15 parts of Dysosma Versipellis, 19 parts of Nidus Vespae, 30 parts of Cortex Phellodendri, 6 parts of Fructus Bruceae, 19 parts of Fructus Cnidii, 7 parts of Radix Glycyrrhizae, 19 parts of Flos Lonicerae, 19 parts of Folium Isatidis, 13 parts of Radix Semiaquilegiae, and 1500 parts of 75% ethanol.

3. The traditional Chinese medicine composition according to claim 1, wherein the traditional Chinese medicine composition is a tincture.

4. A preparation method of the traditional Chinese medicine composition according to claim 1, comprises the following steps: (1) soaking 13-17 parts of Dysosma Versipellis, 17-21 parts of Nidus Vespae, 4-8 parts of Fructus Bruceae, 17-21 parts of Flos Lonicerae, and 11-15 parts of Radix Semiaquilegiae into 95% ethanol for 8-12 days, taking a first supernatant of soaked solution, extracting using the ethanol, and concentrating the first supernatant at 60-70° C., 0.02-0.05 MPa to obtain an ethanol extract concentrate; (2) adding 1400-2500 parts of ionized water to: 36-40 parts of Rhizoma Smilacis Glabrae, 17-21 parts of Sophora Flavescens, 28-32 parts of Herba Hedyotidis Diffusae, 28-32 parts of Herba Taraxaci, 28-32 parts of Cortex Phellodendri, 17-21 parts of Fructus Cnidii, 5-9 parts of Radix Glycyrrhizae, and 17-21 parts of Folium Isatidis, heating and boiling, and taking the filtrate to obtain a water extract; and (3) mixing the ethanol extract concentrate with the water extract, filtering out a second supernatant of the mixed solution, and adding 30% ethanol into the second supernatant to get 700-800 parts of the traditional Chinese medicine composition.

5. The preparation method according to claim 4, comprising the following steps: (1) soaking 15 parts of Dysosma Versipellis, 19 parts of Nidus Vespae, 6 parts of Fructus Bruceae, 19 parts of Flos Lonicerae, and 13 parts of Radix Semiaquilegiae in 95% ethanol for 10 days, taking the a first supernatant of the soaked solution, extracting using ethanol, and concentrating the first supernatant at 65° C., 0.04 MPa to obtain an ethanol extract concentrate; (2) adding 1920 parts of ionized water to 38 parts of Rhizoma Smilacis Glabrae, 19 parts of Sophora Flavescens, 30 parts of Herba Hedyotidis Diffusae, 30 parts of Herba Taraxaci, 30 parts of Cortex Phellodendri, 19 parts of Fructus Cnidii, 7 parts of Radix Glycyrrhizae, and 19 parts of Folium Isatidis, heating and boiling, and taking out the filtrate to obtain a water extract; and (3) mixing the ethanol extract concentrate with the water extract, filtering out a second supernatant, and adding 30% ethanol to get 750 parts of the traditional Chinese medicine composition.

6. The method according to claim 4, wherein the specific gravity of the traditional Chinese medicine composition is measured by a Baume's specific gravity meter.

7. The preparation method according to claim 4, wherein the specific gravity of the prepared traditional Chinese medicine composition is 0.90-1.12.

8. A method of preventing and/or treating condyloma acuminatum comprising administering to a patient the traditional Chinese medicine composition according to claim 1.

9. A method of preventing and/or treating cervical precancerous lesions comprising administering to a patient the traditional Chinese medicine composition according to claim 1.

10. A method of preventing and/or treating skin tinea comprising administering to a patient the traditional Chinese medicine composition according to claim 1.

11. The traditional Chinese medicine composition according to claim 2, wherein the traditional Chinese medicine composition is a tincture.

12. A method of preventing and/or treating condyloma acuminatum comprising administering to a patient the traditional Chinese medicine composition according to claim 2.

13. A method of preventing and/or treating cervical precancerous lesions comprising administering to a patient the traditional Chinese medicine composition according to claim 2.

14. A method of preventing and/or treating skin tinea comprising administering to a patient the traditional Chinese medicine composition according to claim 2.

Description

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0044] The present invention is further illustrated by the following examples, but it should be understood that the scope of the invention is not limited by the specific embodiments.

Example 1 Composition and Preparation Method of Traditional Chinese Medicine Composition

Composition:

[0045] 38 parts by weight of Rhizoma Smilacis Glabrae, 19 parts of Sophora Flavescens, 30 parts of Herba Hedyotidis Diffusae, 30 parts of Herba Taraxaci, 15 parts of Dysosma Versipellis, 19 parts of Nidus Vespae, 30 parts of Cortex Phellodendri, 6 parts of Fructus Bruceae, 19 parts of Fructus Cnidii, Radix 7 parts of Glycyrrhizae, 19 parts of Flos Lonicerae, 19 parts of Folium Isatidis, 13 parts of Radix Semiaquilegiae, 1500 parts of 75% ethanol.

[0046] The preparation method is as follows.

[0047] (1) soaking 15 parts of Dysosma Versipellis, 19 parts of Nidus Vespae, 6 parts of Fructus Bruceae, 19 parts of Flos Lonicerae, and 13 parts of Radix semiaquilegiae in 95% ethanol for 10 days, taking a first supernatant of the above soaked solution. Then the first supernatant is extracted using ethanol, and then the first supernatant is concentrated at 65° C., 0.04 MPa, to obtain an ethanol extract concentrate.

[0048] (2) adding 1920 parts of ionized water to 38 parts of Rhizoma Smilacis Glabrae, 19 parts Sophora Flavescens, 30 parts Herba Hedyotidis Diffusae, 30 parts Herba Taraxaci, 30 parts of Cortex Phellodendri, 19 parts Fructus Cnidii, 7 parts of Radix Glycyrrhizae, and 19 parts of Folium Isatidis, heating and boiling, and taking out the filtrate to obtain a water extract.

[0049] (3) mixing the ethanol extract concentrate with the water extract, filtering out a second supernatant from the above mixed solution, and adding 30% ethanol to get 750 parts of the composition.

[0050] The prepared Chinese medicine composition is measured to by a Baume's specific gravity meter obtain a specific gravity of 0.968.

Example 2 Inhibition of Human Papillomavirus Nucleic Acid Replication In Vitro by the Traditional Chinese Medicine Composition of the Present Application

[0051] First, 30 samples of genital condyloma acuminatum are collected, and Human papillomavirus (HPV) nucleic acid DNA after homogenization is collected.

[0052] Secondly, the traditional Chinese medicine composition prepared in Example 1 is selected, and the composition is mixed with water to make diluted solutions with concentration of 5%, 2.5%, 1.25%, 0.625%, and 0.3125%. The samples are immersed in the solution for 34 hours and 58 hours respectively. The test temperature is 25° C. Saline is used as a negative control.

[0053] The Conclusion is as follows:

[0054] When immersed for 34 hours: no nucleic acid was detected when the concentration of the traditional Chinese medicine composition of the present invention 2.5%; 6.67% of the nucleic acid was detected at the concentration of 1.25%; 50% of the nucleic acid was detected at the concentration of 0.625%; 93.33% of the nucleic acid was detected at the concentration of 0.3125%;

[0055] When immersed for 58 hours: no nucleic acid was detected when the concentration of the traditional Chinese medicine composition of the present invention 1.25%; 26.67% of the nucleic acid was detected at the concentration of 0.625%; and 96.673% of the nucleic acid was detected at the concentration of 0.3125%;

[0056] Saline control group: 100% of nucleic acid was detected after 34 hours and 58 hours.

[0057] It is proved that the traditional Chinese medicine composition of the present embodiment can inhibit the replication of human papillomavirus nucleic acid in vitro, effectively destroy the viral DNA, has the ability to kill the virus, and the inhibition of HPV is strengthened with the prolongation of time and the increase of concentration, The minimum effective concentration is 1.25%.

Example 3 the Traditional Chinese Medicine Composition of the Present Application Treats Condyloma acuminatum

[0058] 1. Firstly, 18-65 years old male and female genital condyloma acuminatum cases were chosen: single wart diameter <0.5 cm, wart numbers <5 (wart mass diameter <1 cm), the wart was located in the external or perianal part. The cases were divided into the test group and the control group, 30 persons each, the test group used the traditional Chinese medicine composition prepared in Example 1 of the present invention, and the control group used the imiquimod cream.

[0059] The following patients are excluded: skin disease patients with impact assessment; severe liver and kidney dysfunction; those who were allergic to imiquimod cream; those who received viral therapy.

2. Test Method:

[0060] Test group: The pharmaceutical composition prepared in Example 1 of the present invention was taken out with a cotton swab and applied to the warts and the surrounding 2 cm area. 2 times/day, continuous use for 3 days, stopped for 4 days, 7 days for a course of treatment. If the wart was not completely detached after the end of a course of treatment, a second course of treatment could be used for up to four courses of treatment.

[0061] Control group: 5% imiquimod cream was applied to the warts and the surrounding 2 cm area with a cotton swab three times a week, every other day for 8 weeks.

[0062] The two groups of patients were re-examined every 2 weeks for 8 weeks. The curative effect was observed and samples were collected for viral marker detection by PQ-PCR.

3. Results

[0063] The subjects were actually completed in 60 cases, 30 cases in the test group, and 30 cases in the control group.

3.1 Complete Shedding Rate of Warts in the First Course of Treatment

[0064]

TABLE-US-00001 TABLE 1 Comparison of complete shedding rate of warts in the first course of treatment of two different drugs Effective number of Invalid number of people in the people in the first course of first course of treatment treatment Total Test group 26 4 30 Control group 18 12 30 Note: χ.sup.2 = 5.455 p = 0.020

3.2 Virus Negatve Detection

[0065]

TABLE-US-00002 TABLE 2 Comparison of the negative rate of HPV virus markers after treatment with two drugs Trans-negative number Invalid number Total Test group 28 2 30 Control group 20 10 30 Note: χ.sup.2 = 6.670 p = 0.010

[0066] 3.3: Recurrence Statistics

TABLE-US-00003 TABLE 3 Comparison of recurrence rate after treatment with two drugs Number of Number of recurrence non-recurrence Total Test group 1 27 28 Control group 5 15 20 Note: χ.sup.2 = 4.898 p = 0.027

[0067] Conclusion: After the first course of treatment with different drug treatments in the test group and the control group, the complete shedding rate of warts, the recurrence rate of cases and the negative rate of HPV virus markers are counted. The comparison results showed that the traditional Chinese medicine composition prepared in Example 1 of the present invention has a remarkable effect on treating condyloma acuminata, and the course of treatment is short (7 days), and the HPV virus markers have a high negative rate and a low recurrence rate.

Example 4 the Traditional Chinese Medicine Composition of the Present Application Acts on Cervical HPV Infection and HPV-Induced Cervical Intraepithelial Neoplasia

[0068] 1. After the confirmation from the hospital outpatients, the test from liquid-based ultrathin (TCT), colposcopy and histopathological examination, the pathological results were confirmed as one of the following conditions: cervical HPV-positive, cervical intraepithelial neoplasia (CIN1, CIN2).

2 Treatment Methods

[0069] All of them were treated in non-menstrual period. The sterilized cotton balls with tail line were used to take 1˜2 ml of the traditional Chinese medicine composition prepared in Example 1 of the present invention, which was applied to the cervix, and was taken out by the patient for about 1-3 hours, 1 time/d, used 3d for a while, rest for 4d, and used 4 courses of the treatment. Reviewed after deactivation.

3 Efficacy Criteria

3.1 HPV Infection

[0070] (1) Cure: HPV DNA review results were negative.

[0071] (2) Invalid: HPV DNA review results were positive.

3.2 CIN Cure:

[0072] (1) Both TCT and HPV tests were negative. Colposcopy, it was also valid if there was no abnormality under the microscope or confirmed by biopsy to confirm the absence of CIN.

[0073] (2) Invalid: pathological diagnosis after biopsy CIN1 or CIN2 remained unchanged.

[0074] 3.3 The χ2 test was used for statistical analysis. P<0.05 was considered statistically significant.

4. Results

[0075]

TABLE-US-00004 TABLE 4 Traditional Chinese medicine composition of the present invention acts on cervical HPV infection and cervical intraepithelial neoplasia Diseases Number of cases Valid Invalid Cervical HPV positive 34 21 13 C1N1 7 6 1 C1N2 4 3 1

[0076] Conclusion: The traditional Chinese medicine composition prepared in Example 1 of the present invention has significant effects on cervical HPV infection and HPV-induced cervical intraepithelial neoplasia.

Example 5 the Traditional Chinese Medicine Composition of the Present Application Acts on Pathogens of the Skin Tinea

[0077] The two most common pathogens, Trichophyton rubrum and Candida albicans, were conducted pre-experiments of the external drug sensitivity by using M-38 A2 (skin sputum) and M27-A3 (yeast) micro-liquid dilution method developed by CLSI experiment.

1 Test Strains

[0078] The pre-experimental strains include 1 strain of Trichophyton rubrum (BMU-08583) and 1 strain of Candida albicans (BMU-05260).

2 External Drug Sensitivity Test Method

[0079] Trichophyton rubrum and Trichophyton rubrum were measured by the M38-A2 micro-liquid dilution method established by CLSI; Candida albicans was measured by the M27-A3 micro-liquid dilution method established by CLSI.

3 Result Judgment

[0080] The results were interpreted in accordance with the standards established by CLSI. Only when the growth control grew well, the test was considered successful and the results could be interpreted.

4 Results

[0081] 4.1 Pre-Experimental Results for Trichophyton rubrum and Candida albicans

[0082] The minimum inhibitory concentration (MIC) of the traditional Chinese medicine composition prepared in Example 1 of the present invention against Candida albicans was 1:8; the MIC against Trichophyton rubrum was 1:128. The MIC of the traditional Chinese medicine composition prepared in Example 1 of the present invention for removing the ethanol stock solution against Candida albicans was 1:8; the MIC for the Trichophyton rubrum was 1:128. The pre-experimental results of the external drug sensitivity of the traditional Chinese medicine composition prepared in Example 1 of the present invention were shown in Table 5.

TABLE-US-00005 TABLE 5 Results of external drug sensitivity test of traditional Chinese medicine composition of the present invention Traditional Chinese Traditional Chinese medicine composition medicine composition containing ethanol without ethanol Strains MIC MIC Candida albicans (1) 1:8  1:8  Trichophyton rubrum (1) 1:128 1:128

[0083] The experimental results suggested that the traditional Chinese medicine composition of the present invention has certain antifungal activity in vitro, especially for the Trichophyton rubrum, and the MIC is 1:128 (that is, the result of diluting the original solution of the Chinese medicine composition by 8 times). It also has antibacterial activity against Candida albicans, and the MIC is 1:8 (i.e., the result of diluting the original solution of the Chinese medicine composition by 4 times). The results showed that the traditional Chinese medicine composition containing and without the ethanol solution has no effect on the results of the external drug sensitivity test, indicating that the ethanol has no antifungal activity, and the true antifungal activity comes from the traditional Chinese medicine ingredients in the traditional Chinese medicine composition of the present invention.

4.2 Results of External Antibacterial Activity Against Trichophyton rubrum and Toe (Finger) Trichophyton

[0084] The MIC of the traditional Chinese medicine composition prepared in Example 1 of the present invention against Trichophyton rubrum was 1:64 to 1:256; the MIC of toe (finger) Trichophyton was 1:64 to 1:128. The consistency of the results of the two tests per strain was very good. The results of external antibacterial activity against Trichophyton rubrum and toe (finger) Trichophyton were shown in Table 6.

TABLE-US-00006 TABLE 6 Results of external antibacterial activity of the traditional Chinese medicine composition against dermatophytes Strains 1:32 1:64 1:128 1:256 1:512 Trichophyton rubrum 10 0 4 4 2 0 toe (finger) Trichophyton 10 0 7 3 0 0 Total 20 0 11 7 2 0

[0085] The traditional Chinese medicine composition prepared in Example 1 of the present invention has obvious antifungal activity to Trichophyton rubrum and Toe (finger) Trichophyton, and the MIC of Trichophyton rubrum is 1:64-1:256. That is, the stock solution of the traditional Chinese medicine composition of the present invention has an antifungal effect when diluted to 7 to 9 times; the MIC of the toe (referring to) Trichophyton is 1:64 to 1:128, that is, the stock solution of the traditional Chinese medicine composition of the present application has an antifungal effect when diluted to 7 to 8 times.

Example 6

Composition

[0086] Rhizoma Smilacis Glabrae 38 parts, Sophora Flavescens 19 parts, Herba Hedyotidis Diffusae 30 parts, Herba Taraxaci 30 parts, Dysosma Versipellis 15 parts, Nidus Vespae 19 parts, Cortex Phellodendri 30 parts, Fructus Bruceae 6 parts, Fructus Cnidii 19 parts, Radix Glycyrrhizae 7 parts, Flos Lonicerae 19 parts, Folium Isatidis 19 parts, Radix Semiaquilegiae 13 parts, Bassia hyssopifolia 12 parts, 75% ethanol 1500 parts.

Preparation Method

[0087] (1) soaking 15 parts of Dysosma Versipellis, 19 parts of Nidus Vespae, 6 parts of Fructus Bruceae, 19 parts of Flos Lonicerae, and 13 parts of Radix semiaquilegiae in 95% ethanol for 10 days, taking out a first supernatant, ethanol-extracting the first supernatant, and concentrating the first supernatant at 65° C., 0.04 MPa to obtain an ethanol extract concentrate.

[0088] (2) adding 1920 parts of ionized water into 38 parts of Rhizoma Smilacis Glabrae, 19 parts Sophora Flavescens, 30 parts Herba Hedyotidis Diffusae, 12 parts of Bassia hyssopifolia, 30 parts Herba Taraxaci, 30 parts of Cortex Phellodendri, 19 parts Fructus Cnidii, 7 parts of Radix Glycyrrhizae, and 19 parts of Folium Isatidis, heating and boiling, and taking out the filtrate to obtain a water extract; and

[0089] (3) mixing the ethanol extract concentrate with the water extract, filtering out a second supernatant, and adding 30% ethanol into the second supernatant to get 750 parts of the.

The Traditional Chinese Medicine Composition of the Present Invention Acts on Cervical HPV Infection and HPV-Induced Cervical Intraepithelial Neoplasia

[0090] 1. After the confirmation from the hospital outpatients, the test from liquid-based ultrathin (TCT), colposcopy and histopathological examination, the pathological results were confirmed as one of the following conditions: cervical HPV-positive, cervical intraepithelial neoplasia (CIN1, CIN2).

2 Treatment Methods

[0091] All of them were treated in non-menstrual period. The sterilized cotton balls with tail line were used to take 1˜2 ml of the traditional Chinese medicine composition prepared in Example 1 of the present invention, which was applied to the cervix, and was taken out by the patient for about 1-3 hours, 1 time/d, used 3d for a while, rest for 4d, and used 4 courses of the treatment. Reviewed after deactivation.

3 Efficacy Criteria

3. 1 HPV Infection

[0092] (1) Cure: HPV DNA review results were negative.

[0093] (2) Invalid: HPV DNA review results were positive.

3.2 CIN Cure:

[0094] (1) Both TCT and HPV tests were negative. Colposcopy, it was also valid if there was no abnormality under the microscope or confirmed by biopsy to confirm the absence of CIN.

[0095] (2) Invalid: pathological diagnosis after biopsy CIN1 or CIN2 remained unchanged.

[0096] 3.3 The χ2 test was used for statistical analysis. P<0.05 was considered statistically significant.

4. Results

[0097]

TABLE-US-00007 TABLE 7 Traditional Chinese medicine composition of the present invention acts on cervical HPV infection and cervical intraepithelial neoplasia Diseases Number of cases Valid Invalid Cervical HPV positive 34 30 4 C1N1 7 7 0 C1N2 4 4 0

[0098] Conclusion: The traditional Chinese medicine composition prepared in Example 6 of the present invention has significant effects on cervical HPV infection and HPV-induced cervical intraepithelial neoplasia.

[0099] The experimental effect of the present embodiment is obviously superior to the experimental effect of the composition of Example 1. Compared with Example 1, the other conditions are exactly the same, only the Bassia hyssopifolia is added, it can be seen that Bassia hyssopifolia has the effect of treating cervical HPV infection and HPV-induced cervical intraepithelial neoplasia.

Example 7

Composition

[0100] Rhizoma Smilacis Glabrae 38 parts, Sophora Flavescens 19 parts, Herba Hedyotidis Diffusae 30 parts, Herba Taraxaci 30 parts, Dysosma Versipellis 15 parts, Nidus Vespae 19 parts, Cortex Phellodendri 30 parts, Fructus Bruceae 6 parts, Fructus Cnidii 19 parts, Radix Glycyrrhizae 7 parts, Flos Lonicerae 19 parts, Folium Isatidis 19 parts, Radix Semiaquilegiae 13 parts, Fauriella 18 parts, 75% ethanol 1500 parts.

Preparation Method

[0101] (1) soaking 15 parts of Dysosma Versipellis, 19 parts of Nidus Vespae, 6 parts of Fructus Bruceae, 19 parts of Flos Lonicerae, 13 parts of Radix semiaquilegiae and 18 parts of Fauriella in 95% ethanol for 10 days, taking the supernatant, and ethanol extracting and concentrating the supernatant at 65° C., 0.04 MPa to obtain an ethanol extract concentrate;

[0102] (2) adding 1920 parts of ionized water into 38 parts of Rhizoma Smilacis Glabrae, 19 parts Sophora Flavescens, 30 parts Herba Hedyotidis Diffusae, 30 parts Herba Taraxaci, 30 parts of Cortex Phellodendri, 19 parts Fructus Cnidii, 7 parts of Radix Glycyrrhizae, and 19 parts of Folium Isatidis, heating and boiling, and taking the filtrate to obtain a water extract; and

[0103] (3) mixing the ethanol extract concentrate with the water extract, filtering out a second supernatant, and adding 30% ethanol into the second supernatant to get 750 parts of traditional Chinese medicine composition.

[0104] The traditional Chinese medicine composition of the present embodiment acts on pathogens of the skin tinea.

[0105] The two most common pathogens, Trichophyton rubrum and Candida albicans, were conducted pre-experiments of the external drug sensitivity by using M-38 A2 (skin sputum) and M27-A3 (yeast) micro-liquid dilution method developed by CLSI experiment.

1 Test Strains

[0106] The pre-experimental strains include 1 strain of Trichophyton rubrum (BMU-08583) and 1 strain of Candida albicans (BMU-05260).

2 External Drug Sensitivity Test Method

[0107] Trichophyton rubrum and Trichophyton rubrum were measured by the M38-A2 micro-liquid dilution method established by CLSI; Candida albicans was measured by the M27-A3 micro-liquid dilution method established by CLSI.

3 Result Judgment

[0108] The results were interpreted in accordance with the standards established by CLSI. Only when the growth control grew well, the test was considered successful and the results could be interpreted.

4 Results

[0109] 4.1 Pre-Experimental Results for Trichophyton rubrum and Candida albicans

[0110] The minimum inhibitory concentration (MIC) of the traditional Chinese medicine composition prepared in Example 6 of the present invention against Candida albicans was 1:64; the MIC against Trichophyton rubrum was 1:256. The MIC of the traditional Chinese medicine composition prepared in Example 1 of the present invention for removing the ethanol stock solution against Candida albicans was 1:64; the MIC for the Trichophyton rubrum was 1:256. The pre-experimental results of the external drug sensitivity of the traditional Chinese medicine composition prepared in Example 1 of the present invention were shown in Table 8.

TABLE-US-00008 TABLE 8 Results of external drug sensitivity test of traditional Chinese medicine composition of the present invention Traditional Chinese Traditional Chinese medicine composition medicine composition containing ethanol without ethanol Strains MIC MIC Candida albicans (1) 1:64  1:64  Trichophyton rubrum (1) 1:256 1:256

[0111] The experimental results suggested that the traditional Chinese medicine composition of the present invention has certain antifungal activity in vitro, especially for the Trichophyton rubrum, and the MIC is 1:256. It also has antibacterial activity against Candida albicans, and the MIC is 1:64. The results showed that the traditional Chinese medicine composition containing and without the ethanol solution has no effect on the results of the external drug sensitivity test, indicating that the ethanol has no antifungal activity, and the true antifungal activity comes from the traditional Chinese medicine ingredients in the traditional Chinese medicine composition of the present invention.

[0112] The experimental effect of the present embodiment is obviously superior to the experimental effect of the composition of Example 1. Compared with Example 1, the other conditions are exactly the same, only the fauriella is added, it can be seen that the fauriella has the effect of killing Trichophyton rubrum and Candida albicans.

Comparative Example 1

Composition

[0113] Rhizoma Smilacis Glabrae in 38 parts, Sophora Flavescens in 19 parts, Herba Hedyotidis Diffusae in 30 parts, Herba Taraxaci in 30 parts, Dysosma Versipellis in 15 parts, Nidus Vespae in 19 parts, Cortex Phellodendri in 30 parts, Fructus Bruceae in 6 parts, Fructus Cnidii in 19 parts, Radix Glycyrrhizae in 7 parts, Flos Lonicerae in 19 parts, Folium Isatidis in 19 parts, and 75% ethanol in 1500 parts.

Preparation Method

[0114] (1) soaking 15 parts of Dysosma Versipellis, 19 parts of Nidus Vespae, 6 parts of Fructus Bruceae, and 19 parts of Flos Lonicerae in 95% ethanol for 10 days, taking out a first supernatant, and ethanol-extracting the first supernatant, and concentrating the supernatant at 65° C., 0.04 MPa to obtain an ethanol extract concentrate;

[0115] (2) adding 1920 parts of ionized water into 38 parts of Rhizoma Smilacis Glabrae, 19 parts Sophora Flavescens, 30 parts Herba Hedyotidis Diffusae, 30 parts Herba Taraxaci, 30 parts of Cortex Phellodendri, 19 parts Fructus Cnidii, 7 parts of Radix Glycyrrhizae, and 19 parts of Folium Isatidis, heating and boiling, and taking the filtrate to obtain a water extract was obtained; and

[0116] (3) mixing the ethanol extract concentrate with the water extract, filtering out a second supernatant, and adding 30% ethanol into the second supernatant to get 750 parts of the traditional Chinese medicine composition.

The Traditional Chinese Medicine Composition of the Present Invention Acts on Cervical HPV Infection and HPV-Induced Cervical Intraepithelial Neoplasia

[0117] 1. After the confirmation from the hospital outpatients, the test from liquid-based ultrathin (TCT), colposcopy and histopathological examination, the pathological results were confirmed as one of the following conditions: cervical HPV-positive, cervical intraepithelial neoplasia (CIN1, CIN2).

2 Treatment Methods

[0118] All of them were treated in non-menstrual period. The sterilized cotton balls with tail line were used to take 1˜2 ml of the traditional Chinese medicine composition prepared in Example 1 of the present invention, which was applied to the cervix, and was taken out by the patient for about 1-3 hours, 1 time/d, used 3d for a while, rest for 4d, and used 4 courses of the treatment. Reviewed after deactivation.

3 Efficacy Criteria

3.1 HPV Infection

[0119] (1) Cure: HPV DNA review results were negative.

[0120] (2) Invalid: HPV DNA review results were positive.

3.2 CIN Cure:

[0121] (1) Both TCT and HPV tests were negative. Colposcopy, it was also valid if there was no abnormality under the microscope or confirmed by biopsy to confirm the absence of CIN.

[0122] (2) Invalid: pathological diagnosis after biopsy CIN1 or CIN2 remained unchanged.

[0123] 3.3 The χ2 test was used for statistical analysis. P<0.05 was considered statistically significant.

4. Results

[0124]

TABLE-US-00009 TABLE 9 Traditional Chinese medicine composition of the present invention acts on cervical HPV infection and cervical intraepithelial neoplasia Diseases Number of cases Valid Invalid Cervical HPV positive 34 18 16 C1N1 7 4 3 C1N2 4 2 2

[0125] Conclusion: The traditional Chinese medicine composition prepared in Example 7 of the present invention has significant treatment effects on cervical HPV infection and HPV-induced cervical intraepithelial neoplasia.

[0126] The experimental effect of the present embodiment is obviously lower than the experimental effect of the composition of Example 1. Compared with Example 1, the other conditions are exactly the same, only the radix semiaquilegiae is not added. It can be seen that radix semiaquilegiae has the effect of treating cervical HPV infection and HPV-induced cervical intraepithelial neoplasia.

[0127] The foregoing descriptions of specific embodiments of the present invention are presented for purposes of illustration and description. It is not intended to be exhaustive or to limit the invention to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. The exemplary embodiments were chosen and described in order to explain certain principles of the invention and their practical application, to thereby enable others skilled in the art to make and utilize various exemplary embodiments of the present invention, as well as various alternatives and modifications thereof. The invention is to be limited only by the claims provided below and equivalents thereof.