At the beginning, HUST-China set up Mr. Tony project with the exact goal of truly make contributions beneficial to the world, and strive to be responsible for people around the world. It means that at the end of completion of the project, it is important to broaden the impact of the project on society and humanity. This is not only instructive to the long-term development of our project and contribution to society, but also useful to the spread of science in the whole society with our own strength.

We tend to change the society's attitude towards hair dyeing and perming via organizing Brave Color Challenge in worldwide. And the widespread of our experience and knowledge is deepened through education in all age. At the same time, our team give out our own strength through social volunteering. Additionally, we develop our product into a complete business plan which software and hardware are included. We also shared our software design on the internet via open source.

1 Brave color challenge -- attention to social prejudice

Figure 1: Brave color challenge

At the beginning of the project, we carried out a wide range of surveys via interviews, questionnaires to inquire consumers, entrepreneurs, stakeholders and different identities, and found that the domestic people may have a certain degree of bias on the style of the exaggerated permed or dyed hair. Therefore, we designed a reversible bio-perm dyeing system that fades instantly, and selected plant dyes to mitigate its harm to health and environment. However, during project surveys and field visits, we found that while most people were interested in Mr. Tony's products with some trust and expectation, the designated product didn't actually eliminate people's biases.

We looked up the relevant information comprehensively and found that prejudice is essentially a negative understanding and attitude to people and things that bulit up from objective facts, just like stereotypes. Prejudice is not easy to be eliminated in a short period of time, and it often requires years of subtle influence to do so. But we wouldn't give up just because it's time-consuming, and HUST-China decided to do its best to do its part in the field of perming and dyeing to eliminate prejudice.

In addition, we also found that the most terrible prejudice is often the prejudice against themselves. Long-term Influenced under growth and living environment, many people become suspicious of themselves after being prejudiced by others. Susan Bealey, M.D. of the University of British Columbia in Canada and health coach, says it's not uncommon, or even quite normal, for perfect and capable people to think they can't. These "no" misconceptions are biases that are embedded by others, often from families where people grew up, and may be judged from a conservative perspective by family members. And the elimination of this prejudice often rely on self-contained self-confidence and a strong heart, telling themselves to be brave and not care about what others think. It is in this way that HUST-China launched the Brave Color Challenge around the world, calling on people to eliminate prejudice and be brave enough to be themselves.

The slogan of Brave Color Challenge is to be brave to find the own self, and draws on the relay model of the Ice Bucket Challenge to expand the project's impact. We allow challengers to change their hair color via Photoshop, eliminating challengers' concerns about hair health and social attitudes, skipping tedious thoughts and selection processes, and just appreciating their hairstyle after dyeing their hair. We not only provided hair color advice for the challenger, more importantly, we aimed to let the challenger find a more beautiful self, and encourage them to be brave take steps to embrace their own identity. After the challenge, many challengers told us that they did challenge themselves and found self-confidence. HUST-China is proud to contribute to the elimination of self-prejudice.

In addition, we have made the benefits of natural dyes widely understood to challengers and the public through knowledge cards in the hope of enhancing public understanding of this potentially large but low market share industry.

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Brave-color-challenge

Challenge process: The Brave color Challenge draws on the relay model of the Ice Bucket Challenge to expand the project's impact by naming the next challenger.

The Challenge process comes below:

1 Select the hair color for the next challenger via our self-constructed program (For detailed information , please refer to software part),

2 The Challenger would use Photoshop or other image-fixing software to change the hair color of their selfie to the color which the previous challenger have chosen, referring to the guidance provided by us

3 Challenger would post her selfie before and after recoloring and the hair color selected for the next challenger on the social website??and @the next challenger.

Figure 2: Brave color challenge

**Interview with Challengers:**After challenge, we have also interviewed a number of challengers, who said that they did have some bias and trouble about hair dyeing. Through this challenge, they found their adoring hair dye color, and they would also try to use less harmful natural perm and dye to explore their more beautiful image. After we introduced our projects to challengers, they really look forward to the reversible and healthy feature of our products.

**Knowledge Cards:**We launched the challenge poster and added the design of the knowledge card afterwards to promote the challenge while spreading the health benefits of natural dyes to the public. In addition, we call on the public to pay attention to the importance of environmental protection.

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2 Education

Education and public outreach are important guidelines for our team. In the past year, we have carried out a series of educational practices mainly aimed at students. In the process of communicating with many schools and institutions, we took the advantages of university platform, and conveyed the team's name and knowledge about synthetic biology to a wider audience. Our audience mainly includes pupils, secondary school students, college students and those who love life science. Our educational activities not only include traditional offline knowledge lectures, communication meetings and road shows, but also offer courses in popular online media, providing online education resources for public, which is a highlight for our team.

Primary School

Figure 3: Primary School

We always believe that the spread of science will not be limited by age. As the first stop of our education program, we came to the Primary School affiliated to Huazhong University of Science and Technology. Together with other scientific research teams and individuals of Huazhong University of Science and Technology, we have carried out a series of interesting science courses called "Dream Class" in the hope that pupils can establish their dreams in class. In this course, we designed an interesting biology Mini Game to tell the children about the basic and interesting biology knowledge, and also introduced the past projects of the HUST-China team to the children in popular language. Unexpectedly, the pupils not only understood the story, but also made wild suggestions to our project.

High School

Figure 4: High School

The middle school period is a critical spot for determining personal professional preferences. In order to increase the influence of iGEM and popularize the knowledge of synthetic biology, our team organized a number of educational activities for middle school students. This educational activity covers middle school students of different backgrounds. With the vivid content of the course, we rooted curiosity of biology in middle school students' hearts.

Universities

Figure 5: Universities

The university itself is a diversified learning platform, for our university students to popularize science provides a lot of convenience. In the education of college students, we rely on the institution of the school to show them synthetic biology and iGEM in a more official and professional image. At the same time, we also attach importance to the training of new team members. Since the beginning of this year, we have inherited the spirit and tradition of iGEM predecessors, and cultivated a group of brand new iGEM team members. In addition, we also actively publicized our humanistic practice overview and gains to the public, so that more people could see how large-scale practical activities are scientifically organized and carried out.

HUST-China team has undertaken part of the "Qiming Courses" to enable students of many other majors to understand synthetic biology-related knowledge, as long as hair dyeing experience activities.

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Universities

Qiming College is an institution established by Huazhong University of Science and Technology (HUST) to cultivate top-notch innovative talents. Qiming College has launched a series of "Qiming Courses" as the interdisciplinary learning platform. Through communication with college, we applied for the qualification to set up the course.

As a representative of basic subject education, the HUST-China team has undertaken part of the "Qiming Courses" to enable students of many other majors to understand synthetic biology-related knowledge. Around synthetic biology, we introduced the basic background knowledge of basic biochemistry and molecular biology.

Figure 6: Our team members are talking about biochemistry related knowledge

In addition, with this platform, we also promoted iGEM: the situation of our team and the basic overview of the 2021 project to students. Closely related to the content of the project, the students were taught the construction of engineered cells and the scientific knowledge of hair dyeing and perming. In the course, we provided hair dyeing experience activities. We provided hair and a variety of dyes for the students . Students could DIY their own hair color and styling, so as to have a deeper understanding of the creativity and feasibility of our project.

Figure 7: Students DIY their hair styling

Figure 8: Student is sharing his work

In other Qiming courses, we specially offered courses related to tie-dyeing craftsmanship, teaching students to dye handicrafts with natural pigments. At the same time, taking this as an opportunity, we highlighted the significance of the engineered cells constructed by our project, and demonstrated the environmental protection and health of green and natural pigment synthesis. With this course, we once took the concept of the project deeply into the hearts of the students and greatly promoted the creativity of the team.

Figure 9: Students show the crafts they made in class

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Figure 10: In-team education

Every year we will provide months of in-team education. The coverage of the member education is very broad, not only includes the necessary theoretical knowledge such as biochemistry, molecular biology and synthetic biology.

Bilibili open course

Figure 11: Bilibili open course

This year, we innovatively launched online courses. With the current trend of online course resource learning becoming stronger, we uploaded a series of online courses on the Bilibili website, which are very popular among Chinese youth groups. The course content covers a wide range, including not only a variety of basic theoretical knowledge, but also a full range of application software teaching, aiming to train the audience to become a qualified iGEM player through the course.

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Bilibili-open-course

Affected by the epidemic, we chose to train our team members through online open courses at Bilibili. Our team has selected some excellent players to teach online courses. The contents of the open course include biochemistry, molecular biology, literature retrieval and so on, In addition, we have also conducted brief training for software such as PS,AE,snapgene and other software to better participate in the competition.

Figure 12: Explanation on the use of snapgene

In these courses, we also give a systematic and detailed introduction to the iGEM competition. Our team members explained in detail the considerations to be considered in designing a project and the steps for project design. In order to better find the genetic information and protein information we need, we have special team members to explain the use and search of databases such as NCBI,KEGG. During the training of the team members, we made a comprehensive recording of the course, uploaded the video to bilibili, the largest video website in China, and optimized its content, in order to help students master basic literature retrieval knowledge and skills. the use of some commonly used software, as well as the explanation of some professional knowledge. We hope that through the online open course, we can let you master more and more comprehensive knowledge, publicize our project with the help of the Internet, at the same time, let more people know about synthetic biology and IGEM events, cultivate students' interest in synthetic biology and increase their knowledge about it.

Figure 13: Explanation of experimental principle

Open courses in partnership with UCAS-China IGEM HUST-China offers courses for team members every year, and in order to better inherit and carry forward the courses, we have recorded some open courses on software and basic knowledge and decided to continue to expand accordingly. However, with the attitude of learning from each other and making progress together, we have decided to work with UCAS-China to continue to expand our courses in more comprehensive areas of knowledge. At present, the plan is in progress.

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3 Social public welfare and influence

Donation of plant dye handicrafts

Figure 14: Donation of plant dye handicrafts

In July 2021, we participated in an education-aiding public welfare activity in Lianhua Village, Pingxiang , Jiangxi Province for 21 days. An investigation about autism in Jiangxi and several commonweal courses including bandhnu, popularization of synthetic biology, etc were applied to students from Xiaoshui Village, Chetian Village, Shanxia Village and Lianhua Village. During the popularization course, a vivid and lucid drama was used to reveal how synthetic biology works and benefits the whole society. From the investigation, we found that there was 29.8% of them suffering from severe autism, which was inextricably related to relationship with parents and friends, while whether their parents were working outside of town had no influence on them. To promote bandhnu and improve the autism and loneliness of the students, we established a bandhnu course towards all grades and specialized one of them. Colourful and various types of clothes, bags and scarves were created from them and displayed in a T show during the farewell party. This public welfare activity expanded their horizon, exposed them to professional knowledge and techniques, enhanced their confidence and did deserve all our effort.

Popularization of synthetic biology

Figure 15: Popularization of synthetic biology

Investigation of autism

Figure 16: Distribution status of suburb children's autism

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Investigation-of-autism

Self-evaluation scale: 'Children Solitude Scale' of S. R. Asher, 1984 was used in the investigation by cluster sampling adopted to students from grade 3 to 6. Giving out 141 questionnaires, getting feedback from all 141, 100% of response rate. After eliminating invalid questionnaires (too regular the answers or with too much deletion) by surveyors manually, we've got 131 effective questionnaires, 92.9% of efficiency. The whole process was anonymous and be granted from subjects of their own will. Under instruction of surveyors, subjects filled the questionnaires on-site. The questionnaires include control point scale and age, gender, number of friends, living standard, status of parents' vocation, and some other demographic indicators.

The result is divided into two parts.

A. Distribution status of suburb children's autism. We have subjects of 74 male (56.5%) and 57 female (43.5%), average age of 10.34±1.32. 45.8% of them seemed to have slight or medium autism while another 29.8% suffering from severe autism. (see Fig1)

Fig1. Distribution status of suburb children's autism

Figure 17: Distribution status of suburb children's autism

B. Distribution feature of suburb children's autism. We found the academic status, number of friends and relationship with parents are related to autism degree (P<0.05) while having nothing to do with status of parents' working out of town (P>0.05). Less the friends, higher the score in autism test. Worse the relationship with parents, worse the autism.

Tab2. Distribution feature of suburb children's autism

Demographic indicators		Total	Status of autism			F	P
		Non-autism	Slight/ Medium	Severe
Academic status	Excellent	27	11(40.7)	11(40.7)	5(18.5)	2	0
	Good	29	8(27.6)	15(51.7)	6(20.7)
	Medium	45	9(20.0)	23(51.1)	13(28.9)
	Mediocre	19	3(18.5)	6(31.6)	10(52.6)
	Work hard	11	1(9.1)	5(45.5)	5(45.5)
Number of friends	None	1	0	0	1(100.0)	-2	0
	1-2	18	3(16.7)	10(55.6)	5(27.8)
	3-5	31	4(12.9)	12(38.7)	15(48.4)
	6-10	35	12(34.4)	15(42.9)	8(22.9)
	>10	46	13(28.3)	23(50.0)	10(21.7)
Status of parents' working out of town	Both	43	11(25.6)	17(39.5)	15(34.9)	-1	0
	Father	48	8(16.7)	26(54.2)	14(29.2)
	Mother	11	3(27.3)	4(36.4)	4(36.4)
	Neither	29	10(34.5)	13(44.8)	6(20.7)
Relationship with parents	Unhappy	9	1(11.1)	6(66.7)	2(22.2)	-2	0
	Not bad	15	1(6.7)	9(60.0)	5(33.3)
	Lovely	57	18(31.6)	20(35.1)	19(33.3)

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Bandhnu course

Figure 18: Bandhnu course

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Bandhnu-course

On the basis of project Mr. Tony, we established a bandhnu course of interest every Monday, Wednesday and Friday and a specialized one in Class one, Grade six. We bought pigment, enhancer, white T-shirts, hats and scarves. We explained the history of bandhnu and modern techniques, which could increase students' manipulative ability and creativity during application. In the farewell party at the end of the education-aiding course, T-show was held to encourage students daring to show off their product and personality. Through the charm of colour, students enhanced their relationship with each other and obviously, removed some of the lonliness.

photo	description
	Dyeing in the bandhnu course
	Displaying bandhnu product from one of the students
	Laughter in their own bandhnu clothes on the T show
	Group photo of students with their own bandhnu product and joy
	'Models' awaiting to going on the stage

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Bioethics week debate

Figure 19: Bioethics week debate

With the invitation of the HZAU-China team, HUST-China travelled to Huazhong Agricultural University to participate in the Bioethics Week. Both sides launched a friendship debate --- ' artificial intelligence / humans are more likely to lead to human destruction '. After the fierce debate between the two sides, the questioning of the audience, the comments of the judges and the sharing of points of views, the audience had a new understanding of life ethics. We do not want human civilization to really go to the day of destruction, but we continue to imagine how it may be destroyed. So we should eliminate and avoid errors, and then move forward more cautiously. It is beyond dispute that through this event, we can be more cautious and also more courageous in the face of scientific progress and assume the responsibility of life science practitioners.

Beijing voluntary lecture

Figure 20: Beijing voluntary lecture

In order to pay attention to the popularization and rapid development of synthetic biology, HUST-China team responded to the call of CAU-China to join in voluntary lectures on synthetic biology for all college students in Beijing community. This voluntary lecture, faced with college students in Beijing, talked a lot about the history of synthetic biology, basic knowledge of life science, brief introduction and prospect of synthetic biology, and introduction of iGEM events, which not only expanded the influence of iGEM events, but also comprehensively popularized synthetic biology. In addition, each team carried on the project display in turn, really let the students combine with the actual application to consolidate the basic knowledge of synthetic biology. We also arranged interesting knowledge answers and questions part, which narrowed the gap between students, and comprehensively answered the questions of students.

Figure 21: Beijing voluntary lecture

After the lecture, many listeners said that this event made them have some preliminary understanding of the principles and applications of synthetic biology. The projects introduced by the lecturers were very meaningful and promising, and they said that they would continue to further understand and learn relevant knowledge in the future.

4 Commercial products

Software:

1. Based on the algorithm framework of YOLOv5, we customize a set of data sets and data labels on the micronucleus count of vicia faba root tips, which includes hundreds of typical chromatin and micronucleus images. Based on this, a model is trained to count the micronucleus of vicia faba root tips under the experimental conditions of this project, and then determine the toxicity of various dyes. We opened the download of this model on GITHUB, and introduced the self-defined process in MODEL ( if it is only used for the identification of micronucleus ) to ensure that each user can train a counting model suitable for its personal experimental conditions.

2. According to the results of hair dyeing experiments, we developed a set of color prediction algorithms, and developed a small program named Dye Your Hair-Mr. Tony HUST on WeChat. This widget returns the expected hair color results based on the type of pigment you submit and the dyeing time. The selection of single / two pigments is currently supported. Maintenance will continue to be updated.

For detailed information , please refer to software part

Hardware:

1. We have designed a portable and easy-to-use fermenter whose fermentation chamber and reaction chamber are integrated into one device. After fermentation, the working enzyme can pass through the filter and produce pigments in the reaction chamber only by inverting the bottle, which will make it more convenient for the production and storage of pigments and provide convenience for users.

2. We also designed a dye comb to cooperate with the use of dye in the fermentor. This tool will enable users to customize the entire process from pigment production to hair dyeing.

For detailed information , please refer to hardware part

Business plan:

In order to make a further progress of the following projects and exaggerate the influence of Mr. Tony brand, we have also formed a business plan to continue to promote a healthy, humanized and reversible biological perm and dye system.

5 Open source project sharing

1. MCN counter : Based on the algorithm framework of YOLOv5, we customize a set of data sets and data labels on the micronucleus count of vicia faba root tips, which includes hundreds of typical chromatin and micronucleus images. Based on this, a model is trained to count the micronucleus of vicia faba root tips under the experimental conditions of this project, and then determine the toxicity of various dyes. This greatly liberates the manpower and the heavy reading work will be done by the computer. We opened the download of this model on GITHUB, and introduced the self-defined process in MODEL ( if it is only used for the identification of micronucleus ) to ensure that each user can train a counting model suitable for its personal experimental conditions.

2. Dyeing predictor : We built several BP neural networks in python3 + tensorflow + keras environment to predict the dyeing results according to the type, concentration and time of pigment. If you ' ve read the relevant parts of MODEL in detail, you ' ll realize that this network is actually for all fiber dyeing - pigment attachment systems. Therefore, you can completely customize your own prediction model according to this network structure. This model contributes more in the mathematical part of determining data labels and activation functions of each layer. You can learn more in the relevant parts of MODEL.

Marvellous! Human practice conducted by your team not only served as a guide for the project, but also contributed to the world by disseminating synthetic biology knowledge and conveying positive energy!! Now, let's have a look at Modeling part!