Human social practice means thoughtful engagement with people at all levels of the world as they use synthetic biology to solve real-world problems. It has great significance to our project design and laboratory work. We divided human practice activities into three aspects: consultation of professionals, interview of patients, and communication between teams. In terms of professionals, we consulted dietitians and doctors, who provided detailed opinions on our project and resolved many of our concerns during the course of the project. About patients, we kept following up the patient interviews and went to the patients' homes to have in-depth and full communication with them. On inter-team cooperation, we have conducted in-depth discussions and cooperation with the University's affiliated team on laboratory safety issues.

1. Project origin

-Through the investigation, a frequently appearing term attracted our attention: glucosinolates
-This caused us to think deeply: Does glucosinolate have the ability to prevent cancer and protect the intestines? ? ?

2. Data retrieval

-We conducted some data searches and filled in a lot of background knowledge in order to come up with a more formal and academic research topic.
Research: 
Glucosinolates are a class of compounds found in cruciferous vegetables such as broccoli, cauliflower, cabbage, Brussels sprouts, and kale. When these vegetables are chewed or chopped, an enzyme called myrosinase converts glucosinolates into bioactive compounds, including isothiocyanates (ITCs) and indole-3-carbinol (I3C). These bioactive compounds have attracted significant scientific interest due to their potential health benefits, including their possible role in cancer prevention and intestinal protection.
Glucosinolates, a class of compounds found in cruciferous vegetables, have been studied for their potential ability to prevent cancer and protect the intestinal tract. Here are some key points based on existing research:

Cancer Prevention:
Glucosinolates are converted into bioactive compounds, such as isothiocyanates (ITCs), when cruciferous vegetables are chewed or chopped.
ITCs have demonstrated anticancer properties in laboratory and animal studies. They can inhibit cancer cell growth, induce apoptosis (programmed cell death) in cancer cells, and inhibit tumor formation.
ITCs may exert their effects through various mechanisms, including modulation of detoxification enzymes, inhibition of angiogenesis (formation of new blood vessels to support tumor growth), and suppression of inflammation.

Intestinal Tract Protection:
The intestinal tract plays a crucial role in overall health, and maintaining its integrity is important for preventing various diseases, including colorectal cancer.
Glucosinolates and their breakdown products, such as ITCs and indole-3-carbinol (I3C), have shown potential protective effects on the intestinal tract.
These compounds may help reduce inflammation, protect against oxidative stress, and promote a healthy balance of gut microbiota, which are important factors in maintaining intestinal health.

Some studies have suggested that I3C can modulate signaling pathways involved in the regulation of intestinal barrier function, potentially enhancing intestinal protection.
It's important to note that while there is promising evidence from laboratory and animal studies, more research is needed to fully understand the effects of glucosinolates on cancer prevention and intestinal health in humans. Clinical trials and epidemiological studies are necessary to evaluate the impact of these compounds on cancer risk reduction and intestinal protection in human populations.Furthermore, individual variations in metabolism, genetic factors, and dietary factors may influence the effectiveness of glucosinolates in different individuals. Therefore, it is essential to consider these factors when interpreting the potential benefits of glucosinolates.To stay updated on the latest scientific findings, it is recommended to consult reputable scientific journals, databases, and consult with medical professionals or experts in the field of oncology or nutrition. They can provide the most current and accurate information regarding the potential role of glucosinolates in cancer prevention and intestinal protection.


https://platform.almanhal.com/Files/Articles/92512
Rameeh, V. (2015). Glucosinolates and their important biological and anticancer effects: A Review. Jordan Journal of Agricultural Sciences, 11(1), 1-13.
-Glucosinolates and their breakdown products have long been recognized for their fungicidal, bacteriocidal, nematocidal, and allelopathic properties and have recently attracted intense research interest because of their cancer chemoprotective attributes. 
-Glucosinolate derivatives stop cancer by destroying cancer cells, and they also suppress genes that create new blood vessels, which support tumor growth and spread.
-These organic compounds also reduce the carcinogenic effects of many environmental toxins by boosting the expression of detoxifying enzymes. 

Li, Z. Y., Wang, Y., Shen, W. T., & Zhou, P. (2012). Content determination of benzyl glucosinolate and anti–cancer activity of its hydrolysis product in Carica papaya L. Asian Pacific journal of tropical medicine, 5(3), 231-233.
-A number of studies support the fact that a thioglycoside constituent, benzyl glucosinolate (BG) exists in all tissues except the mature pulp of Carica papaya[1]. In the catalysis of myrosinase, BG can be hydrolyzed into benzyl isothiocyanate (BITC), a compound that has cancer-preventive and anti-cancer activities[2,3]. 
-Carica papaya L. is a perennial evergreen herbaceous plant belonging to Family Caricaceae and Genus Carica, which has a short growth period and its fruit is of great nutritive and health-care value [4]. Further research and exploitation of high-added-value products from papaya will benefit the development of the papaya industry. In this study, Cultivar “Sunrise Solo” papaya was used as the plant material. The contents of BG in the papaya pulps and seeds
Summary at the end this stage: 
-It's important to note that while there is promising evidence from laboratory and animal studies, more research is needed to fully understand the effects of glucosinolates on cancer prevention and intestinal health in humans. Clinical trials and epidemiological studies are necessary to evaluate the impact of these compounds on cancer risk reduction and intestinal protection in human populations. Furthermore, individual variations in metabolism, genetic factors, and dietary factors may influence the effectiveness of glucosinolates in different individuals. 
-We raised a further question: Can glucosinolates from the Brassicaceae be used in the treatment of intestinal cancer?

3. Nutritionist project verification

We confirmed that glucosinolate can be used and effective, and we also communicated with a nutritionist through social media. In addition, we were also able to establish connections with a nutritionist, who hopes to provide us with long-term professional guidance and help.

4. Patient interviews

We interviewed many patients and their families and found that the diet and recommendations given by doctors do include a lot of broccoli
- So we found experts on the Internet or in real life who could answer our research topic
-Interviewing experts. 
- We put some questions together and made some questions

Email content: 
Subject: Request for Expert Opinion on the Potential of Glucosinolates in Cruciferous Family for Bowel Cancer Treatment

Dear Professor XXXXXX,

I hope this email finds you in good health and high spirits. My name is Cici and I am a member of Thinker China, and we are a groups of high school students that are doing a genetic genernerring competition. I am reaching out to you today because I greatly admire your expertise and research contributions in the field of (), particularly your work in ().

I am currently exploring the research topic of utilizing glucosinolates found in the cruciferous family of vegetables as a potential treatment for bowel cancer. Given your extensive knowledge and experience in the field, I would greatly appreciate your expert opinion on the subject matter.

The cruciferous family, including vegetables such as broccoli, cabbage, and cauliflower, is known to contain high levels of glucosinolates. These compounds have been linked to various health benefits, including potential anticancer properties. My research aims to investigate the mechanisms by which glucosinolates could potentially be used in the treatment of bowel cancer, as well as their efficacy and safety.

I have conducted an extensive literature review to gather relevant information on this topic, and while I have come across some promising findings, I believe that your insights would greatly enrich my understanding. Therefore, I kindly request a few minutes of your valuable time to seek your professional opinion regarding the following points:

Are you aware of any recent studies or developments regarding the potential use of glucosinolates in the treatment of bowel cancer?
What are your thoughts on the efficacy and safety of utilizing glucosinolates for bowel cancer treatment?
In your expert opinion, what are the major challenges or limitations that need to be addressed in this research area?
Are there any specific areas or aspects of this research topic that you believe require further investigation?

Please note that any information or insights you can provide, including relevant publications, would be immensely beneficial to my research. Moreover, I am open to any suggestions you may have regarding additional sources or experts who could contribute to my study.

I understand that you are incredibly busy, but your expert opinion would be of immense value to me. I genuinely appreciate your time and consideration in reviewing this email. Thank you in advance for your assistance.

Please feel free to suggest a convenient time and method for us to discuss this topic further, either via email or through a phone call. I am eager to learn from your expertise and insights. I have attached my contact details below for your convenience.

Thank you once again for your attention and anticipated support. I look forward to hearing from you.

Sincerely,
Cici and Thinker China 
IGEM

5. Online project discussion

Format: on online/phone chat

Key point: Consider the possible development branches of the project and the feasibility of each branch, and determine the preliminary direction
We brainstormed online, discussing the every branch of the project that could develop and their feasibility. We also studied current colorectal cancer treatment options to understand their strengths and weaknesses, and identified issues that our program needed to improve and solve. After in-depth discussion, we preliminarily determined the direction of the project.
After viewing the data, we understand the reason that broccoli can fight cancer: according to the National Cancer Institute, the secret of cruciferous vegetables' anti-cancer ability is that they are rich in high concentrations of thioglucoside.
Sulforaphane is a kind of isothiocyanate, obtained by the hydrolysis of glucosinlates (Glu) by the myrosinase enzyme in the plant. The molecular formula is C6H11NOS2.
We learned in detail why broccoli fights cancer, and learned that glucosinolates are its main anticancer components, and myrosinase is the key to the conversion of glucosinolates into anticancer components sulforaphane.
- Core gene: myrosinase
The conversion of glucosinolate to sulforaphane requires the hydrolysis of myrosinase, which is involved in the plant's defense against herbivores. However, mammals do not express myrosinase, and the conversion effect of the host microbiota to glucosinolate is weak and insufficient to induce anticancer effects. To achieve this, we designed an engineered strain that can express myrosinase and use myrosinase to produce sulforaphane from the cruciferous vegetable diet.

1. Interview stakeholders

Interview stakeholders
Doctors and patients are the core beneficiaries of our programs. Before visiting, we will carefully design a series of questions to ensure that we can obtain their valuable feedback and comments to further optimize our project.

1) Questionnaire design

(1) Doctors
- What do you think about the use of engineered bacteria as an adjunct cancer treatment strategy?
What do you think are the advantages and disadvantages of this strategy compared to existing prevention methods?
- What challenges do you think patients might face when using such products?
(2) Patients and their families
- Have you heard of the use of engineered bacteria as a strategy for cancer adjuvant therapy before?
- How familiar are you with how cancer can be treated through diet and lifestyle?
- What do you think about using engineered bacteria to prevent colorectal cancer?
- What do you care most about when using this product?
- What factors do you think will influence your decision to use this engineered bacteria?

2) Interview

After in-depth interviews, doctors reported that our proposed method is innovative compared with traditional treatments, and sulforaphane is also effective in fighting cancer. At the same time, the patients and their families also actively expressed their attitude: as long as the safety can be ensured, they are very willing to try new products related to this, but the patients' families and patients still have certain concerns about the use of genetically engineered bacteria in human beings.

2. Nutritionist sharing

The dietitian shared with us a copy of the article that curcumin can treat colorectal cancer. This piqued our interest. After reviewing the data, we found that curcumin in ginger is also an effective anti-cancer substance, but because of its strong taste, many people can not accept it, so we decided to carry out a new study to try to extract curcumin from ginger.
Core genes: Diketide-CoA synthase (DCS), Curcumin synthase (CURS)

 -We decided to use the ferulic acid pathway to synthesize curcumin

1. Security issues

Since our products are directly applied to the human body, safety is a primary consideration. In the development process, we paid special attention to the selection of suicide genes, which need to have a significant effect on E. coli, while ensuring that there is no harm to human cells. We face some difficulties in finding the right suicidal gene and are not sure how to proceed with further research.

Therefore, we decided to turn to experts for help and interviewed the experienced Dr. Li. In our in-depth conversation with Professor Yang, we gained valuable advice: we can consider using SRRz cutting genes, which target cell walls. This opens up new directions and hopes for our project research.
Core gene: SRRz cutting gene
Linkage gene composed of phage lysozyme (transglycosidase) gene R and gene RZ. The product of the R gene is a water-soluble transglycosylase, a peptidoglycan that breaks down cell walls. The product of the RZ gene is an endopeptidase that splits between the oligosaccharides of peptidoglycan and cross-links between the peptidoglycan and the outer membrane of the cell wall. The function of S gene products is to change the permeability of the plasma membrane and form a porous structure on the plasma membrane, so that the enzymes produced by R and RZ genes can cross the plasma membrane and reach the cell wall, thus acting on the cell wall, causing the cell wall to break and the release of intracellular substances. Both R and RZ gene products function to degrade cell walls.
- We decided to cut the gene by using SRRz

2. Product conversion

Designing inducible promoters that can effectively induce the activation of suicide genes has been a difficult problem for us, and despite great efforts, we still have not been able to find the ideal inducible promoter. Moreover, patients and their families also have certain concerns about the use of genetically engineered bacteria in human bodies.

Figure 1. Offline discussion

Figure 2. Communicate with Teacher Liu
We made a deliberate decision to shift the scope of our products from in vivo to in vitro. Under the guidance of the teacher, we selected a temperature-controlled promoter to induce the expression of the suicide gene. This not only further enhance the biosafety of our products, but the previously selected lytic genes also help improve the efficiency of our engineered bacteria. We firmly believe that this transition will make our products more stable, secure and provide a solid foundation for future applications."
- Select items for in vitro product use

1. Patient acceptance

Key point: Whether to accept our products
In the trial phase of our product, patient feedback is our primary concern. Their opinions and feelings will directly influence the future improvement of our products.
However, considering the psychological problems of the patient, we found the family members of the patient interviewed before to conduct a second interview. In the face of the new product we introduced, families made it clear that they felt more comfortable and at ease with this in vitro application strategy compared to the engineered bacteria used directly in the human body. This external use gives them a more intuitive and accessible treatment option.

2. Doctor interview

Visiting the doctor with our new product. The doctor praised our new product.
1) The product does not interact directly with human cells, thereby reducing potential risks;
2) In vitro application is more convenient for monitoring and adjustment;
3) Provides patients with a relatively easy and more acceptable treatment

3. Hardware design

As our strategy shifted, adapting products from in vivo to in vitro applications, we faced new challenges. In order to ensure that users can easily and quickly enjoy our products, our team decided to develop a custom hardware device after discussion. This device can be precisely matched to our product characteristics, and with user friendliness in mind, it can provide users with smooth and simple operation.
"Hardware according to design drawings"

4、Better lab work

Before conducting the experiment, we evaluated all possible safety issues. We believe that despite safety training, our team members may still forget the correct procedure during the course of the experiment. In the process of communicating with other teams, we learned that the RDFZ-China team had similar problems to us, so our two teams decided to cooperate in this area. We started with an online meeting to provide management ideas to each other on laboratory safety.

After the communication, we set up a group on social media (WeChat) to plan the specific contents of the cooperation in detail. First of all, we will paste detailed instrument operation process and prompt slogans on the instrument. Secondly, we will paste two-dimensional code on the experimental instrument. After scanning, you can view cartoons about the operation steps and operational risks of the experimental instrument, so that students can have a more intuitive understanding of the operation process. In this process, our slogan and QR code resources will be shared in the group (WeChat) at any time to improve the efficiency of problem solving.

Figure 3. Detailed instrument operation procedures

Figure 1. slogan