Sampling, culturing and analyzing method for pustular acne and application thereof

Abstract

The invention belongs to the technical field of skin analysis and nursing, and discloses a pustular acne sampling, culturing and analyzing method and application thereof. Comprising the steps that volunteer face pustular acne and normal skin photographing is conducted, volunteer face pustular acne and normal skin part microorganism sampling is conducted, microorganism samples are cultured, and treatment is conducted through image J and a pH meter. Based on skin surface pH value regulation and control and zymophyte characteristic analysis, the composition has the biological activity effects of resisting acnes, repairing skin barriers and regulating skin micro-ecology.

Description

Technical Field

[0001] The invention belongs to the technical field of skin analysis and care, and particularly relates to a pustular acne sampling, culture and analysis method and application thereof. Background Art

[0002] Acne is a common skin disease caused by blocked hair follicles, excessive secretion of sebaceous glands, inflammatory response and imbalance of microbial flora. Traditional studies have shown that the pH value of acne skin is usually high, showing an alkaline environment, which may promote the growth of harmful bacteria such as Propionibacterium and aggravate acne symptoms. [1,2] To this end, treatment often involves acid scrubbing, which aims to inhibit the activity of Propionibacterium by lowering the pH of the skin. [3] Pustular acne are small bumps and inflammatory lesions that develop on the skin through excess oil and dead skin cells clogging the pores. Pustular acne are inflammatory lesions that contain yellow fluid or pus in the center.

[0003] However, after in-depth research on acne skin microorganisms, our new findings show that the pH value on the surface of pustular acne skin is actually acidic and there are a large number of fermentative bacteria. This is in stark contrast to the traditional view, indicating that the microbial environment of acne sites is different from what was previously known. The fermentative flora also contains a variety of probiotics, and the presence of these beneficial microorganisms may play a positive role in regulating the skin microecology. [4] . Summary of the invention

[0004] In order to overcome the shortcomings of the prior art, the present invention provides a pustular acne sampling, culture and analysis method and its application, which provides a new idea for the effective treatment of acne by judging the pH value of the skin surface of pustular acne and combining the characteristics of fermentation bacteria.

[0005] The above object of the present invention is achieved through the following technical scheme: a method for sampling, culturing and analyzing pustular acne, comprising the following steps:

[0006] 1. Take photos of the volunteers' facial pustular acne and normal skin;

[0007] 2. Microbial sampling of volunteers' facial pustular acne and normal skin;

[0008] 3. Cultivate the microbial sample in step 2;

[0009] 4. Process the results in step 3 using image J and a pH meter.

[0010] Furthermore, the step 1 is specifically as follows: before conducting the experiment, ensure that the acne patient is without makeup to avoid external contamination interfering with the results. Use the original camera of the mobile phone to take pictures of the pustular acne part and the normal skin part.

[0011] Furthermore, in step 2, microbial sampling is performed on facial pustular acne and normal skin:

[0012] Use a sterile cotton swab dipped in sterile saline to sample the sampling part (the cotton swab is gently rolled for 3 weeks, 1 ± 0.1 cm 2 ), and the cotton swab with the sample was cut with sterile scissors and placed into a test tube of trypticase soy broth medium (Solarbio, Beijing) for microbial culture.

[0013] Furthermore, the step 3 specifically includes: culturing the microbial sample in step 2:

[0014] (1) Liquid culture medium culture: Cut the cotton swab with the sample into a test tube containing trypticase soy broth medium (Solarbio, Beijing) and 0.5% bromocresol purple indicator and place it in a 37°C incubator for culture. Use a pH meter to measure the pH value every 3 hours, and observe the changes at 0, 3, 6, and 9 hours.

[0015] (2) Solid culture medium culture: The bacterial liquid cultured in liquid culture medium was measured by OD600 with a microplate reader to determine the bacterial count (1x10 6 mL / CFU. Take 20ul 1x10 6 mL / CFU was dropped onto a 9 cm culture dish containing trypticase agar medium (Solarbio, Beijing) and 0.5% bromocresol purple indicator, and evenly spread with a spreader bar. The culture dish was placed in a 37°C incubator for 18 hours.

[0016] Furthermore, the step 4 specifically includes analyzing the experimental results using imageJ and a pH meter.

[0017] Another object of the present invention is to protect the use of the fermented bacteria formed after the above-mentioned pustular acne sampling and culturing in the preparation of drugs for treating pustular acne.

[0018] The beneficial effects of the present invention compared with the prior art are as follows: based on the regulation of the pH value of the skin surface and the analysis of the characteristics of fermented bacteria, the present invention has the biological activity of anti-acne, repairing the skin barrier and regulating the skin microecology; through the application of fermented bacteria, the microecological environment of the skin is improved, thereby effectively reducing the recurrence of acne symptoms. In addition, the metabolites of the fermented bacteria may have biological activity that enhances the natural immunity and anti-inflammatory function of the skin. The non-destructive treatment scheme of the present invention provides a new way to manage acne that is safer and milder. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0020] Figure 1 Pictures of normal and pustular acne of volunteers;

[0021] Figure 2 Normal skin and pustular acne fermentation results chart;

[0022] Figure 3 Results of the proportion of fermentative bacteria in normal skin and pustular acne; DETAILED DESCRIPTION

[0023] The present invention is described in detail below by specific examples, but the protection scope of the present invention is not limited. Unless otherwise specified, the experimental methods used in the present invention are all conventional methods, and the experimental equipment, materials, reagents, etc. used can be obtained from commercial channels.

[0024] Example 1

[0025] Method for sampling, culture and analysis of pustular acne

[0026] S1: Photos of volunteers with pustular acne and normal skin on their faces.

[0027] Before the experiment, make sure the acne patient is without makeup to avoid external contamination interfering with the results. Use the original camera of the mobile phone to take pictures of the acne part and the normal skin part.

[0028] S2: Microbial sampling of volunteers' facial pustular acne and normal skin.

[0029] Use a sterile cotton swab dipped in sterile saline to sample the sampling part (the cotton swab is gently rolled for 3 weeks, about 1cm 2 ), and the cotton swab with the sample was cut with sterile scissors and placed into a test tube of trypticase soy broth medium (Solarbio, Beijing) for microbial culture.

[0030] S3: Cultivate the microbial sample in step 2

[0031] (1) Liquid culture medium culture: Cut the cotton swab with the sample into a test tube containing trypticase soy broth medium (Solarbio, Beijing) and 0.5% bromocresol purple indicator and place it in a 37°C incubator for culture. Use a pH meter to measure the pH value every 3 hours, and observe the changes at 0, 3, 6, and 9 hours.

[0032] (2) Solid culture medium culture: The bacterial liquid cultured in liquid culture medium was measured by OD600 with a microplate reader to determine the bacterial count (1x10 6 mL / CFU. Take 20ul 1x10 6mL / CFU was dropped onto a 9 cm culture dish containing trypticase agar medium (Solarbio, Beijing) and 0.5% bromocresol purple indicator, and evenly spread with a spreader bar. The culture dish was placed in a 37°C incubator for 18 hours.

[0033] S4: Data Analysis

[0034] The results in step 3 were processed using image J and a pH meter.

[0035] Figure 1 Picture of normal skin and acne on a volunteer's face. Pustular acne is an inflammatory lesion with yellow fluid or pus in the center.

[0036] Figure 2 The fermentation results of normal skin and pustular acne are shown in the figure. It can be seen that at 6 hours, pustular acne has obvious fermentation acid production and the pH is acidic. It can be inferred that the pH value on the surface of pustular acne is more acidic than that of normal skin.

[0037] Figure 3 The figure shows the percentage of fermentation bacteria in normal skin and pustular acne. The left picture shows the plate before culture, which is purple; after 18 hours of culture, the color of the plate gradually turns yellow as the fermentation bacteria increase. By comparison, the left picture shows that the number of fermentation bacteria in pustular acne is significantly higher than that in normal skin. The right picture is the result of counting the percentage of fermentation bacteria in the total flora based on Image J software. The percentage of fermentation bacteria in pustular acne is 89.6% ± 10.395, while the percentage of fermentation bacteria in normal skin is 35.5% ± 2.555, which further shows that the number of fermentation bacteria on the surface of pustular acne is significantly higher.

[0038] The above-described embodiments are only preferred embodiments of the present invention, but not all feasible embodiments of the present invention. For those skilled in the art, any obvious changes made thereto without departing from the principles and spirit of the present invention should be considered to be included in the scope of protection of the claims of the present invention.

Claims

1. A method for sampling, culturing and analyzing pustular acne, characterized in that: The following steps are involved: S1. Volunteers took photos of pustular acne and normal skin on their faces; S2. Microbial sampling of volunteers’ facial pustular acne and normal skin; S3. Cultivating the microbial sample in step S2; S4. Process the result in step S3 using image J and a pH meter.

2. The method for sampling, culturing and analyzing pustular acne according to claim 1, characterized in that: The step S1 specifically includes: before conducting the experiment, ensuring that the acne patient is without makeup to avoid external contamination interfering with the results. Using the original camera of the mobile phone to take pictures of the pustular acne part and the normal skin part.

3. The method for sampling, culturing and analyzing pustular acne according to claim 1, characterized in that: The step S2 specifically includes: using a sterile cotton swab dipped in sterile physiological saline to sample the sampling part, and using sterile scissors to cut the cotton swab with the sample into a test tube of trypticase soy broth culture medium (Solarbio, Beijing) for microbial culture.

4. The method for sampling, culturing and analyzing pustular acne according to claim 3, characterized in that: The sampling method is: the cotton swab is gently rolled for 3 weeks to obtain 1±0.1cm 2 of the sample.

5. The method for sampling, culturing and analyzing pustular acne according to claim 3, characterized in that: The trypticase soy broth medium was from Solarbio, Beijing.

6. The method for sampling, culturing and analyzing pustular acne according to claim 1, characterized in that: The step S3 is specifically as follows: Liquid culture medium culture: Cut the cotton swab with the sample into a test tube containing trypticase soy broth medium and 0.5% bromocresol purple indicator and place it in a 37°C incubator for culture. Use a pH meter to measure the pH value every 3 hours and observe the changes at 0, 3, 6 and 9 hours respectively; Solid medium culture: The bacterial liquid cultured in liquid medium was measured by OD600 with a microplate reader to determine the bacterial count of 1x10 6 mL / CFU; take 20ul 1x10 6 mL / CFU was dropped onto a 9 cm culture dish containing trypticase agar medium and 0.5% bromocresol purple indicator, and spread evenly with a spreading stick; the culture dish was placed in a 37°C incubator and cultured for 18 hours.

7. The method for sampling, culturing and analyzing pustular acne according to claim 1, characterized in that: The step S4 specifically includes analyzing the experimental results using imageJ and a pH meter.

8. Use of the fermented bacteria formed after sampling and culturing pustular acne as claimed in claim 1 in preparing a drug for treating pustular acne.

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