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Overview

When choosing the theme of the project, we visited the glasses shop offline after searching the information online. Our team found that eye diseases were more likely to occur in contact lens wearers. To help these people, we decided to design an engineered bacterium that can produce the DMBT 1 protein against Pseudomonas aeruginosa and express the antimicrobial peptide RNase7 and the immunomodulator ST2 to provide a more convenient and healthy glasses maintenance experience.

In the early research and development, we through the questionnaire survey and word cloud, further understand the public for the actual situation of eye care, we found that: although contact lenses have come into our lives, but the public still know how to clean contact lenses is still insufficient, and most people are not willing to tedious correct cleaning steps lead to not effective to clean contact lenses, cause eye inflammation. This deeply touched us and strengthened our desire. We decided to design a glasses box containing the inhibition of Pseudomonas aeruginosa containing DMBT 1 protein glasses fluid. When users use the glasses, it can not only achieve ordinary cleaning effect, but also effectively inhibit eye inflammation.

We received outstanding advice and guidance from several experts whose introduction to the shortcomings of preventive precautions to further recognition of the great need for control. At the same time, their suggestions can also help us to improve the design of experiments, such as the sensitivity of eye medication, protein expression and extraction problems, etc. They suggested that we provided a great help in the course of our experiment and solved the problems we encountered.

In the middle of the study, we went to the south lake park, and introduced our products, their positive positive feedback let us further understand the importance of my project and their social responsibility, more determined we produce more convenient and quick effect good contact lens solution idea, let more people can stay away from the eye disease.

Furthermore, we strive to make our work as inclusive as possible, creating a strong social impact. For example, we drew humorous, easy-to-understand cartoons with complex messages through images and small amounts of words, which allowed us to reach a wider and more diverse audience. In addition to these cartoons, we also engage in a variety of popular science activities to increase the interest of people of different backgrounds and ages. In addition, we went to Tongyu County,a former state-level poverty-stricken county, to donate books and conduct publicity activities to promote the further promotion of synthetic biology to special areas.

The integration of industry, education and research can better solve the problems caused by eye diseases caused by improper contact lens cleaning. Therefore, we conducted a survey of the synthetic biology industry, learning how to commercialize the technology to promote the commercialization of our products themselves. We are committed to making the products no longer armchair, but really make good and inexpensive goods, into the ordinary people's homes, give play to its unique advantages, in order to benefit all mankind. Research on commercialization enables the project to truly meet people's needs, which also represents a "closed cycle"

Set the project content

Glasses shop visit

With the development of technology and lifestyle changes, more and more people have become myopic, resulting in the need to wear glasses to improve their vision. As vision correction devices, these glasses play a key role in helping students see their surroundings more clearly in study and life. And in recent years, with the development of science and technology and the advancement of medical technology, contact lenses, as a new type of vision correction method, are more and more favored by students on campus. Students who once relied on glasses now have the option of wearing contact lenses to correct their vision. The popularity of contact lenses has not only changed the appearance of students, allowing them to have confident smiles and free expressions, but also brought many practical conveniences and comforts. But at the same time, more and more people are suffering from eye diseases.

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Figure 1 Someone who is wearing contact lenses

After searching the information online and communicating through offline visits to the eyeglasses shop, our team found that eye diseases were more likely to occur in the contact lens wearers. So our team decided to find out the root cause of the problem and the appropriate solution

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Figure 2: Group photo after the interview.

After reviewing data and literature, we found that improper lens cleaning can breed a variety of bacteria (mainly Pseudomonas aeruginosa), causingcorneal infection and other eye inflammation under certain conditions. And after wearing contact lenses, eye discomfort, fear of the adverse consequences of contact lenses and choose to wear a large number of people, due to improper wearing of lenses, causing bacteria to breed eye inflammation and other diseases frequently occur, we plan to design an engineering bacteria to make it target the growth of eye fungi after contact lens wearing and prevent eye discomfort. Perceptions of the mechanism by which contact lens wearing makes the cornea susceptible to infection have focused on the bacterium Pseudomonas aeruginosa, which is the main cause of lens-associated microbial keratitis. Then if you want to inhibit the inflammation caused by contact lenses, you can start by inhibiting Pseudomonas aeruginosa.

So our team started from the inhibition of Pseudomonas aeruginosa, and we found that the glycoprotein DMBT1 is the main component responsible for inhibiting the convulsive motor function of Pseudomonas aeruginosa, DMBT1 is a ~340 kDa glycoprotein, originally isolated from saliva, involved in mucosal innate immunity. To solve the inhibitory effect of inhibiting Pseudomonas aeruginosa, and considering the stability of the ocular immune microenvironment, it is necessary to introduce gene antibacterial and immunomodulator modules. The use of RNase7 and ST2 can further inhibit related immune regulation, which can well maintain the stability of the ocular immune microenvironment and inhibit other pathogenic bacteria. We combined the two and biologically based on E. coli, designed an engineered bacteria that can generate DMBT1 protein against Pseudomonas aeruginosa and express the antimicrobial peptide RNase7 and immunomodulator ST2, which is exactly the perfect biosynthetic product we envisioned.

Explore the necessity and significance of the project

Questionnaire

Our project aims to design a contact lens solution that contains engineered bacteria that we plan to synthesize to ensure that contact lenses are clean, safe and play a role in preventing eye diseases. Next, in order to further understand the current eye use of the public and the penetration rate of contact lenses, our team went to Nanhu Park to conduct a questionnaire survey and one-on-one interview. We hope that through this interview, we can better discover the problems people face in real life, make our products better fit people's needs, and further expand the meaning of our projects.

We designed two detailed questionnaires on relevant questions to understand people's awareness of contact lens care, usage habits and product needs.We collected nearly 300 questionnaires across gender, age group and occupational background, which can fully reflect the attitudes and opinions of various groups. In this questionnaire, we mainly investigate five areas we concerned, whether the public know the basic knowledge about eye diseases and experience them, whether they know how to prevent and treat eye inflammation, how often people wear contact lenses, how much do they know about proper cleaning of contact lenses, and how much they demand our products.

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Figure 3: Have you or anyone around you ever worn contact lenses?

Questionnaire 1

A total of 180 valid questionnaires were collected in the questionnaire, and the survey results showed that 80% of the respondents had the experience of wearing contact lenses, but 74.44% of the respondents did not know how to clean contact lenses correctly. It is noteworthy that more than 50% of respondents use multifunctional care solutions for disinfection, and a small number use physical disinfection and hydrogen peroxide disinfection. Only 12.22% used the treatment solution to rub the lenses, and others think that cleaning is more troublesome and only occasionally rubbed, or even not cleaned at all. The survey found that more than half of the people have experienced eye inflammation, and the causes of eye inflammation focus on improper cleaning of contact lenses, rubbing eyes with hands, etc. By analyzing the results of the questionnaire, we found that although contact lenses have entered our lives, the public still has insufficient understanding of how to clean contact lenses, and most people are unwilling to carry out cumbersome and correct cleaning steps, resulting in ineffective cleaning of contact lenses, causing eye inflammation.

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Figure 4: Do you know the correct way to clean contact lenses?
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Figure 5: How do you care for and clean your contact lenses?

Questionnaire 2

We also conducted a questionnaire survey on professional knowledge of medical students in various places, and the questionnaire results showed that more than half of the respondents had knowledge about eye diseases, but 70.37% of the respondents did not understand the methods of eye examination, and 67.9% of the respondents did not know the cause of redness and congestion, itchy or painful eyes. 66.67% of respondents said that they did not know about the treatment of eye inflammation such as antibiotics and hormone therapy, which indicates that there is still considerable room for the popularization of treatment for eye diseases.

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Figure 6:Knowledge of common methods of eye examination
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Figure 7:Understand the possible causes of red, bloodshot, itchy or painful eyes.

Word cloud production

We created a word cloud related to "contact lenses" and "synthetic biology" by using the Google Trends query for keyword heat trends . Research has found that "anti-inflammation" is a frequently searched term, causing widespread public concern. This shows that the problem of eye inflammation caused by the improper use of wearing contact lenses can not be ignored.

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Figure 8 Word cloud

To sum up, we believe that eye diseases have now troubled the current generation to some extent, but most people do not have a special understanding of examination and preventive treatment. The advantage of our glasses case is that it contains a special glasses liquid containing the DMBT1 protein, which has the effect of inhibiting Pseudomonas aeruginosa. When users use this glasses liquid to clean glasses, it can not only achieve ordinary cleaning effect, but also effectively inhibit eye inflammation. It could also solve what the public considers to be the biggest problem right now, which is eye disease suppression. In the current environment where eye diseases are more common, our glasses cases also have good market prospects.

Experimental design and improvement of synthetic biology

Interviews with experts Gong Peiyuan

In order to get more professional suggestions and guidance, we went to the campus hospital to ask the doctor of the ophthalmic clinic for project suggestions. The doctor explained to us the shortcomings of the current preventive measures against pseudomonas aeruginosa. most of the preventive measures is to use chemicals spraying directly, and often use the same type of chemicals may lead to pseudomonas aeruginosa resistance. The bacteria are highly adaptable and may gradually develop resistance to commonly used chemicals, making prevention and treatment more difficult. There are secondary environmental impacts, including spraying with a large number of chemicals, including the potential for pollution of soil, water, and other ecosystems. This led us to further realize the huge demand for prevention and treatment, and also affirmed our core competitiveness of using DMBT 1 protein, our products mainly physically act on aeruginosa pseudomonary cells, to overcome the disadvantages of chemical agents.

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Figure9 Photo in front of the hospital

Doctors also pointed out the sensitivity of eye medication, mainly in terms of microbiota density and side effects on the human body. So our team tried to find an automatic control method, and after reviewing the data, we found that using quorum sensing allows cells to express different proteins according to the concentration of bacteria. Quorum sensing is an important way of cell-to-cell communication, which refers to the phenomenon that individual cells perceive the density of bacteria through the enrichment of a self-synthesized self-inducer molecule. When the concentration of self-inducers reaches a certain threshold as the density of bacteria increases, certain genes are turned on like switches, initiating the expression of a subsequent series of genes. So we plan to use the population-sensing suicide gene circuit to give host bacteria the property of initiating suicide at a certain density to control the bacterial density. We will design a plasmid of threshold regulatory protein, and use a suicide switch to enable E. coli to commit suicide according to the microenvironment after expressing the corresponding protein, so that it can form density periodic oscillations to avoid extreme situations.

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Figure10 Pictures of interviews with experts Gong Peiyuan

Interviews with experts Xie Yanyan

After conducting comprehensive market research and collecting guidance from experts, the JLU-China team embarked on practical experiments in our laboratory to validate our hypotheses. We established a co-culture model involving Pseudomonas aeruginosa and corneal epithelial tissue cells, with the key variable being the presence or absence of DMBT1 protein. Our primary objective was to assess the effectiveness of this protein in preventing eye infections by measuring the degree of infection in the epithelial tissue based on experimental data.In addition to this, we conducted cultivation experiments with Pseudomonas aeruginosa using various concentrations of DMBT1 protein. We meticulously recorded the growth patterns of the bacteria, performed linear regression analyses on the experimental data, and visualized the results to better understand the relationship between DMBT1 protein concentration and bacterial growth.

In order to further improve the inhibitory performance of our protein on aeruginosa pseudomonocytes, we went to Jilin University Hospital again to ask expert physicians for improvement advice. After understanding our project plan and experimental results, the doctor decided that the choice of using the DMBT1 protein to inhibit Pseudomonas aeruginosa was a very creative solution, but in practical application, two key factors need to be considered. The first is the expression and extraction of the protein to ensure the purity and stability of the protein. In addition, the formulation and use of the glasses cleaning solution is also very important to ensure that the protein can fully play the bacteriostatic effect without causing any negative effects on the user's eyes. Moreover, doctors pointed out that the antibacterial drugs on the market have reduced disinfection efficacy when CL care products coexist with glasses cases, and have limited effect on inhibiting pathogenic bacteria such as Pseudomonas aeruginosa.

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Figure11 Photo with expert Xie Yanyan

Inspired by this information, we decided to develop a smart contact lens case that integrates a special eyewear liquid containing the bacteriostatic protein DMBT1 to provide a more convenient and healthy eyewear care experience. The DMBT1 protein will help inhibit the growth of pathogens such as Pseudomonas aeruginosa, providing healthier eye protection. At the same time, we will also introduce a smart glasses case, which can monitor the indicators of the care solution in the box in real time, determine whether the care solution is contaminated and whether it needs to be replaced, so as to further reduce the risk of user infection, protect the quality of glasses and extend the service life. In order to achieve comprehensive and timely feedback, our team also uses computer vision recognition to track the course of eye inflammation, and displays experimental data through a website to achieve all-round eye care. Through this exchange, we deeply felt the professional knowledge and scientific research power of the university hospital, and it is this cooperation that enables academic research to be combined with practical scenarios to bring new innovations and advances in the field of eye care and eyewear care.

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Figure12 Group photo after expert interview

After preliminary experiments, we found that temperature had a significant effect on protein through variable experiments. Through consulting doctor, we learned that protein-toxin HicA mainly functions by binding and cutting RNA, while protein-antitoxin HicB has antagonistic effects with HicA. When HicA: When the proportion of HicB was higher, the cells remained dormant. However, when the temperature rises, the thermosensitive promoter CI857 will cause the transcription of the downstream gene FLP enzyme and bind to the FRT site, remove the HicA gene and change the proportion of HicA: HicB, thus releasing the dormancy state of E. coli and making it grow normally. Given the limited range of temperatures that the human eye can withstand, we decided to further find a temperature balance between the two to maximize the product's function under the right conditions

Public feedback

Park interviews

After conducting a series of experiments, we went to Nanhu Park in Changchun City for a random interview. We first interviewed non-myopic people and learned that they had different understanding degrees and attitudes towards contact lenses. Then we interviewed people who have worn lenses, asked them about their understanding of contact lens cleaning methods and protection products, and surveyed how they clean contact lenses in daily life and whether they will use specific contact lens care solution. Most of the respondents did not know the specific cleaning steps, so in order to take the simplest approach, They also shared with us their experiences of eye disease or discomfort caused by improper cleaning. Finally, we told them about our project and asked if they would like to try this new contact lens solution, which is better at treating inflammation caused by bacteria. We got a variety of responses. Some said they wanted to know more about the ingredients and effects of the product and needed more information to make a decision. Most people say yes because they care about their eye health and want to reduce the risk of eye disease. This interview not only allowed people to further understand the correct way to care for lenses, but also had a new understanding of contact lens care solution. We also deeply realized the importance of cleaning and vision protection when using contact lenses, and further understood the importance of our project and our social responsibility. More firm in our idea of producing more convenient and effective contact lens care solution, so that more people can stay away from eye diseases.

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Figure13、14 South Lake Park interview picture

Publicity and promotion

In previous interviews and surveys, we found that most people did not have a good understanding of synthetic biology. In order to further improve people's understanding of synthetic biology and their understanding of IGEM events, and to expand the influence of our projects, we also held a variety of activities.

Draw comics

Considering the knowledge of synthetic biology and the many professional terms involved in our project itself, it is relatively boring and not easy to be accepted by the public. In order to enhance the visual appeal and attract readers of different ages and cultural backgrounds, we have specially produced a series of images and painted them into cartoons. In this way, comics on the one hand can explain complex synthetic biology concepts, processes, or data through images and a few words, making this information more accessible to the public, on the other hand, comics are often easy to share, and they are associated with vivid images and storylines. This helps to improve the memory and dissemination of propaganda messages.Especially on social media, readers often share interesting or engaging comics, thus expanding the spread of promotional messages.

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Figure 16.17.18.19 Some comic book images are shown

At the same time, these vivid images were also made into relevant cultural and creative peripheral products, such as key chains, clips, etc., and distributed in our follow-up publicity activities, further expanding the influence of our project and synthetic biology.

Joint education in schools

Together with other IGEM teams in the school, we organized a synthetic biology science seminar for all teachers and students. In the lecture, the School of Life Sciences team shared the wonderful world of synthetic biology, the Sino-Japanese Friendship Hospital team brought the wonderful science of iGEM competition, and the team of Basic Medical College introduced the cutting-edge knowledge of microbial therapy. Our team led us to have an in-depth understanding of the application of computer in synthetic biology. At the same time, we also introduced our respective projects through PPT and the broadcast of promotional videos, and explored the infinite possibilities of technology together!

Finally, there was an interactive question-and-answer session, and the audience enthusiastically asked questions about synthetic biology, igem competition, and the team events, and we answered them very patiently. After the activity, we also received dozens of feedback after the lecture. All of them were very eager to participate in this competition and hoped to get the help and guidance from our team.

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Figure 20 The interaction of our team with the audience

In general, this lecture not only brought us wonderful popular science knowledge, but also showed everyone's enthusiasm and innovation. Thank you for the wonderful sharing, and thank the audience for their active participation! We look forward to the future journey of scientific exploration, move forward together and create more miracles!

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Figure 21 Presentation scene pictures

Social responsibility

promotion of Matilda Primary School

Tongyu county former as a state-level counties, because of the long-term drought climate conditions and east alkali xisha soil conditions, low crop yield, drought, wind and other natural disasters caused by production and failure situation commonplace, considering the poverty may somewhat limit children, we donated a lot of popular science manuals and books, and in Changchun xinglong hills mingde elementary school launched a unique synthetic biology preaching: "care burning eyes, spectrum biological mystery". During the synthetic biology lecture, we prepared some simple experimental demonstrations for the students to demonstrate the principles and applications of synthetic biology. Creative manual activities were organized to stimulate students' creativity and imagination, and to guide them to online learning platforms and science websites to learn in-depth about synthetic biology and other related fields. We encourage students to maintain their curiosity and enthusiasm for learning, and to stimulate their interest in and love for science. We hope that this activity can plant seeds in the hearts of the students, who may become future scientists, engineers or medical experts in the future, contributing to the health and development of the society.

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Figure 22 Photos of the students after our donated books

Commercialization

It's a great pleasure to see an exciting breakthrough in the field of synthetic biology! However, this is just a new beginning, and we look forward to the infinite possibilities of the future. We are committed to commercialize technology to achieve wider social interests. Synthetic biology has unlimited potential, and we believe that through commercialization, we can promote revolutionary innovation, provide brand new solutions, promote sustainable development, and safeguard public health

Therefore, we produced a synthetic biology research report, and learned how to commercialize the technology to promote the commercialization of our products themselves and bring the technology to the market to promote the development of eye health. We not only need to meet the needs of the scientific field, but also to ensure the commercial success of this technology, so that more people can benefit.

Through continuous efforts and innovation, we hope to bring more innovation, sustainability and well-being to the society. This is the goal we pursue and the mission of the future.