During our visit to the Jiangsu Garden Expo and Zhejiang Yu Village, our team was surprised to find out that the mine sites had changed dramatically. This sparked our curiosity about the concentration of copper ions in the surrounding soil and water. We dug deep and conducted research of specific resources. We investigated and compared most of the exisiting copper ion detection methods, and realized that it is crucial to develop detection methods of copper ions that are rapid.

Therefore, we decided to create a biological method to accurately and rapidly detect the concentration of copper ions. Through this method, we can better understand the distribution of copper ions in the environment and take the necessary measures to protect the health of soil and water. This innovation will provide an efficient and reliable tool for monitoring copper ion concentration, not only for the Garden Expo and Yu Cun, but also for other fields. We believe that through this invention, we will have a better understanding of the impact of copper ions on the environment." The efficiency and accuracy of this approach will allow us to take more timely measures to protect our natural resources and ecosystems.

The comparison of Garden Expo in the past and now：

The comparison of Yu Cun in the past and now：

In order to understand the public's awareness of copper ions, we conducted a questionnaire survey. By sending out questionnaires and collecting responses from different groups of society, we obtained data from about 700 samples. These data can be visually analyzed in the form such as bar charts, pie charts.

We found that ordinary people's knowledge of copper ions is relatively limited. To fill this knowledge gap, we produced a bilingual copper ion publicity brochure in English and Chinese. This manual aims to educate the public about copper ions and provides concise and clear explanations and examples. Through the release and dissemination of the manual, we hope to increase the public's awareness and understanding of copper ions, so as to attract more people's attention to the copper ion pollution.

Such research and awareness campaigns will help us establish effective communication channels with the public and raise public awareness of copper ion pollution. By increasing public awareness of copper ions pollution, we can further promote and apply our copper ion detection methods to protect the environment and human health.

Understanding the advantages and disadvantages of different detection methods allows us to better evaluate the strengths and applicability of our copper ion detection methods, and continue to improve and optimize our technology to reach the optimal conception of our products. Therefore, during our research, we visited water treatment plants, water testing centers, soil research institutes and wastewater treatment plants. These organizations provided their own methods and techniques, contrasting with our methods.

(1) Investigating in water plants:

We went to investigate in Shangyuanmen Water Treatment Plant. By learning about the copper ion contamination at a specific water plant and how they handle the problem, we can determine if there will be potential consumers for our copper ion sensor. We may face some difficulties in this step, as water treatment plants do not usually have the most prominent standards for heavy metals, and plant managers may question the feasibility of biosensors, arguing that biological methods such as yeast have difficulties surviving and may not have sufficient detection accuracy at low copper ion concentrations in domestic water.

（2）Investigate at a wastewater treatment plant：

We went to the sewage treatment plant to investigate and learned the copper ion pollution level in the water body is still low. However, the wastewater treatment plant provided us with the relevant market, and we learned that industrial wastewater plants often use chemical detection methods with high accuracy and efficiency to detect copper ions. This raises the question of whether our biosensors have higher detection accuracy and can survive in specific environments.

(3) Investigate at Institute of Soil Science:

We went to investigate at the Institute of Soil Science, Chinese Academy of Sciences. We aim to understand the copper ion contamination in the soil and the relevant treatment methods, assessing whether there are potential users of our sensor. They proposed that in places with less copper ions, we need to consider whether the sensitivity of the sensor is high enough, and whether the environment will threaten yeast survival. Based on these considerations, the experiments we are going to do are: what is the detection limit of copper ion concentration and how much copper ion concentration will impact yeast.

In the previous social studies, we learned that there are still many problems and improvements in the practical application of our biosensor. In order to better promote the progress of our project and solve the problems that have been exposed, we went to communicate with professional scientific researchers.

(1) Interview PIs of universities:

After talking to the specific institutions regarding water bodies, we realized the shortcomings of our project. To be more clear about the potential advantages and potentials of the project, we went to consult professional scientific researchers. We first interviewed Professor Zhao of the School of Environmental Sciences at Southeast University and Professor Gao Lei of Marine Science at Nanjing Normal University to find out whether the existing chemical methods are good enough and where is our superiority. The professors believe that biological methods should be more convenient and simple to operate. Chemical instruments, which are complex and expensive, so we can make a breakthrough from this perspective.

(2) Interviewing Teacher Yiming Dong:

At the 10th CCiC Conference, Mr. Dong proposed constructive questions about our project. She asked whether our biosensors were environmentally safe, and inspired us that biological methods for detecting copper ions were more environmentally friendly than chemical methods. From this, we decide to upgrade the hardware we had sketched out earlier. In the first round of hardware DBTL, we wanted to add verkon to quickly dispose of biological waste after each usage.

(3) Interview with Yuxin Huang, Institute of Synthetic Biology, Chinese Academy of Sciences, team leader njtech_China2020 :

We interviewed Ms. Huang Yuxin about the accuracy and detection convenience of our yeast sensor. She provided us with resources (see the notes on the right) and consulted relevant literature. We concluded that to solve the problem of whether the accuracy can be improved, a secondary genetic system, galactose induction system, can be introduced. The background expression level of pCUP1 promoter was decreased. In terms of whether the detection can be more convenient, we learnt that the detected copper ion concentration can be expressed through the olfactory output.

（4）Interviewing Ms Xiaoyu Zhao:

To resolve the challenges to monitor the activity of yeast sensors in the environment in real time, and cumbersome preparation for observing fluorescence intensity using flow cytometry, we went to interview a college student who is researching Raman imaging technology. Ms. Zhao Xiaoyu, an undergraduate at Imperial College, a PhD student at ETH Zurich, an Assistant Vice President at China Starwood Capital, and the Founder and CEO of SuperVision Medicine. We asked about the combination of Raman imaging and yeast technology in our interview. According to her, Raman imaging technology can build up a 3D model of a sample non-invasively, rapidly (about five minutes) and with high precision (250 nm) without pre-treating the sample, and detect the distribution and concentration of drugs in each part of the sample, which can be widely applied. She agreed with our idea of combining it with the yeast biosensor, indicating that in the future our overall project can further improve punctuality and stability.

After the project was further upgraded, we went back to the society again to communicate with people in different fields to discuss whether our project has a development prospect in this field.

(1) Interviewing pharmaceutical companies:

After our team learned about the potential benefits of our sensor through conversations with university teachers and researchers, we designed a hardware that is both inexpensive and easy to operate. In order to understand whether this hardware has market development potential in actual pharmaceutical companies, we interviewed Wang Zhengjun, director of Jiangsu Provincial Chinese Pharmaceutical Research Institute. In the interview, we found that for pharmaceutical companies, yeast growth cycle is longer and can be greatly affected by environmental factors. Therefore, it is not suitable for large-scale, high-speed application required by pharmaceutical companies, with potential biosafety issues. Also, Director Wang also put forward some suggestions for hardware products. First, the threshold value of the yeast sensor should be determined, and the field suitability for applying the yeast sensor should be investigated to avoid excessive influence of the environment on the yeast survival. For our products, hardware can continue to be simplified the designed, for example, focusing on a semi-quantitative evaluation such as "whether the test result is positive" to solve the accuracy problem. Therefore, we started the second round of DBTL in our hardware design, which changed the original method of sending samples to the laboratory to measure fluorescence intensity by flow cytometry to test copper ion concentration into a convenient way of observing transparent circles.

(2) Go to the hospital for investigation:

In order to understand the possible diseases caused by copper ions and whether our yeast sensor has the potential to develop in future medical copper ion detection methods in the human body, we interviewed doctors. These include Yongsheng Wang, director of the Neurology of Jiangsu Provincial Hospital of Traditional Chinese Medicine, and Jing Zhang , a doctoral student of Anhui Provincial Hospital of Traditional Chinese Medicine. Anhui Provincial Traditional Chinese Medicine Hospital has advanced neuromedical technology in China and even the world, so their experience in dealing with diseases caused by copper ions is rich. During the interview, after understanding the copper ion itself and the diseases it can cause, the doctor also agreed with our idea. But at the same time, questions and some suggestions were also raised. The first question is whether the extremely low probability of treatment is worth investing a considerable amount of manpower, material resources, and energy. Of course, our design concept and sensors have potential for development in the medical field, and the new generation of medical researchers also have prospects for the project. In addition, Wang Xudong, the director of the internal medicine department of Drum Tower of Nanjing Hospital, mentioned the possible situation and precautions of copper ion deficiency, as well as protective measures for employees engaging in copper related work. In the interview with Dai Xinzheng, the deputy chief physician of the Department of Hepatobiliary Surgery of Jiangsu Provincial People's Hospital, he introduced to us in detail the Wilson disease that may be caused by copper ions, although the cases are very rare and not a common disease.

Dong Yiming, co-founder and chief scientist of Xinsu Technology, and Professor Liu Jinrong, a senior iGEM instructor from Lanzhou University, put forward constructive questions and suggestions for our project, giving us experience and knowledge related to our team. Miss Dong inspired us to design a hardware that could reduce the survival pressure of yeast in the environment.

Bao Yuhan, an iGEM Human Practices Program Office, commented on our HP work. He emphasized that timing and logic are the keys to succeed in HP work, and cognition and communication with stakeholders are essential. In addition, having a rational view of the project's boundaries and limitations, keeping innovate and seeking for novelty are necessary as well.

After the project presentation, we communicated with the iGEMers from Tianjin University and Nanjing University on each other's topics, from which we drew inspiration for the direction of the following development of our project.

We have also been invited to participate in an offline meetup activity with other 2023 iGEM teams. At the meeting, we exchanged opinions and cooperated with each other, during which we learnt about each others’ experimental progress and communiate our ideas and future plans for HP work.

The suggestions and comments received at this CCiC meetup not only helped us perfect our experimental content, but also provided us with inspiration for our future development.

At the meeting, Nanjing-BioXstem and Nanjing-NFLS from Nanjing, Squirrel CHINA and BGI-MAMMOTH from Beijing, UV-diabeaters from Shanghai, and United team, Bs United China shared topics and experiences of their iGEM projects.

One of the two guests, Bao Yuhan, an iGEM Human Practices Program Officer, provided professional comments and suggestions for each team. Also, he emphasized the essence and judging standards of HP activities to us.

The other guest, Zhang Zhiyu, who has rich experience in HP, shared his perspective that HP activities are about showcasing how our researches affect the world and how the world affects our researches, reminding us the HP details we should pay attention to.

On May 28th, the Nanjing-BioXstem team gathered at the RoSSo International Art Center to participate in the "Global Village" art design, environmental protection, and handicraft activity, as well as the team's ice breaking activity.

Students from all over the world have made their own "Global Village" models, experiencing the charm of nature, raising environmental awareness, also realizing the importance of detecting heavy metal pollutants for environmental protection.

Thank you very much for the support and sponsorship of RoSSo teachers for this event.

On June 7th, Zhiyuan Li taught elementary school students at Shuren Primary School in Nanjing about copper ions. Through this lecture, we realize that both in the field of scientific research and daily life, we need to pay more attention to the subtle changes in the micro world. By revealing the interactions between microorganisms and the environment, we can better face various challenges and find innovative solutions. In the following lectures, we will focus on the importance of copper ions in environmental protection and balance of nature. We introduce the copper ion detection method in a concise and vivid manner and combine it with the role of microorganisms to demonstrate the potential and application prospects of this technology to the audience.

On June 22nd, Zimo Lv, on behalf of the Neuroscience Society, presented our project in the Nanjing Jinling High School.

Some students were curious about the practicality of our research content, so they asked about the impact and application of the topic in daily life. The answer was related to monitoring and treatment of sewage plants, as well as the harm of copper ions to human health. But as time passed, our team also encountered similar problems. With the help of professors, everyone gathered their ideas and came up with answers ranging from maintaining the soil environment to exploring livable planets in outer space. Such a brainstorming session materialized our goals, making the students enthusiastic.

We use official accounts and Twitter to publicize our project, aiming to show the public the process and results of our activities, and to popularize the knowledge of synthetic biology to the public. Through the official accounts, we elaborated on the opening ceremony, topics, and various meet up conferences. We issued a questionnaire to survey the public's understanding of copper ions. We've also covered some HP events and will continue to share them in future events. Through publicity, we hope to let more people know about our research, and pay attention to our team, our project and synthetic biology.