1. Overview

The human gut, home to trillions of microorganisms that form the gut microbiota, plays a crucial role in maintaining overall health. An imbalance in this microbial community, known as gut dysbiosis, can impact digestion, metabolism, immune function, and mental health. (Legan et. al, 2022)

The recent rise in the prevalence rate of diseases related to gut dysbiosis, including Crohn's disease which is chronic and currently incurable, and seemingly unrelated disorders, reveals the prevalence of gut dysbiosis. This motivates our team to find solutions to alleviate gut dysbiosis-related symptoms and improve conditions for patients in the long run.

The issue gained further relevance when several teammates experienced upset stomachs due to increased academic pressure, reduced sleep hours, and reliance on fast food for lunch and dinner. All these matters are prevalent with local high school students. This led us to investigate the prevalence of gastrointestinal disease in China, particularly in Beijing, and we found the situation concerning.

Our team focused on detecting common pathogens through both in vivo and in vitro methods, including strains like B. cereus and S. aureus targeted by 2022 and 2023 BNDS-China projects. Based on the idea of mediating key metabolites involved in Inflammatory Bowel Disease and Major Depressive Disorder, namely butyrate and IAA, we aim to expand the two pathways to apply to more scenarios that may lead to gut dysbiosis. To improve our project's feasibility and applicability, we surveyed Beijing residents.

In July, we interviewed a gastroenterologist about the background concerning gut dysbiosis. Her expertise greatly promoted our understanding of the nature of gut microbiota alterations and their relation to the subhealth dietary structure of modern humans. She also elaborated on the drawbacks of current probiotic products and suggested shifting our focus to alleviating the symptoms of patients in recovery or with mild disease. We were advised to consider the durability and colonization of our probiotics. She also enumerated synthetic biology's great potential in Gastrointestinology application.

Curious about potential alternatives to our probiotic platform and how our product would fare in the eyes of experts in the field, we subsequently interviewed a synthetic biologist about cutting-edge technologies and state-of-the-art designs utilized in pharmaceuticals and gut probiotics.

2. Questionnaire to Beijing Residents

A survey questionnaire constitutes a convenient and effective method within the social sciences for data collection, efficiently facilitating the acquisition of relevant information. To enhance the comprehensiveness of our project, we decided to implement questionnaires at the beginning of our Human Practice initiative.

In the initial phase of questionnaire development, we meticulously complied with the reflection cycle we established. This reflection cycle is composed of three core principles:

1. Reflective: Clarifying our objectives to ensure that our human practice activities remain aligned with the overall goals of the project.

2. Responsible: Emphasizing reliability and care, ensuring that our activities adhere to fundamental requirements of safety, authenticity, and dependability, while taking responsibility for our project, the community, and the field of synthetic biology.

3. Responsive: Actively addressing and resolving issues based on the feedback obtained from the questionnaires.

Taking these elements into account, we precisely defined the objectives of distributing the questionnaires. Our primary aims were to assess the health status of Beijing residents concerning intestinal diseases, evaluate their understanding of the relationship between intestinal disorders and other health issues such as psychological conditions, and obtain their perceptions of existing treatment measures. Additionally, we hope to be clear with the public’s acceptance and opinions regarding our project’s products. These objectives were intended to validate the accuracy of our project’s goals and expectations and to ascertain whether the development efforts of other groups within our project were necessary and appropriately directed. Considering our location in Beijing and recognizing our limitations as high school students in terms of technical expertise and resources, we confined the scope of our questionnaire participants to Beijing residents. To capture a comprehensive understanding of diverse experiences, the distribution of questionnaires was randomly and both Chinese and English versions were prepared.

Upon thorough consideration, we proceeded to formulate the questionnaire items. Excluding the initial question regarding participants’ ages, the questionnaire is divided into three sections. The first section involves questions 2-4, which inquire about participants’ knowledge of health issues such as gut diseases, gut dysbiosis, and the interplay between intestinal diseases and psychological disorders such as depression, as well as the general health situation of their gut. To facilitate participant comprehension and serve as a form of public education, these questions include specific examples (e.g., gut dysbiosis) and explanations of the terminology used. The second section, encompassing questions 5-10, explores whether participants have received treatment for intestinal issues, the specifics of such treatments, and their perceptions of existing treatment options. This section presents various situations and multiple-choice options, supplemented by examples to aid participants with similar experiences in making informed selections. The third section consists of questions 11 and 12, which assess participants’ understanding of the relevant aspects of our project’s products and their acceptance of our product. Following the design of the questionnaire, we promptly distributed it on online social platforms, striving to engage individuals from diverse backgrounds. After one week, we concluded data collection, yielding a total of 246 valid responses.

Subsequently, we commenced the analysis of these samples. The sample has an age distribution predominantly of 18 years and under (40.2%) and 40-65 years (47.6%) (Figure 1). This distribution well represents student and parent groups.

Analysis of the second question revealed that over half of the respondents (54.1%) possess some understanding of gut dysbiosis and related intestinal diseases, whereas less than half (40.2%) comprehend the relationship between intestinal diseases and psychological conditions (Figure 2). These findings indicate a moderate public awareness of intestinal health issues, underscoring the necessity for subsequent educational initiatives, such as public lectures, by our team.

A significant proportion of participants (65.0%) reported experiencing intestinal discomfort or being diagnosed with intestinal diseases, highlighting the prevalence and severity of intestinal health problems among the surveyed Beijing residents. However, among those who have symptoms but not disease, only a minority (24.6%) sought hospital treatment, while the majority (53.8%) opted for self-management (Figure 3). This suggests a potential lack of urgency in seeking medical attention among Beijing residents, although it is important to consider that many participants may not belong to age groups with a high incidence of severe intestinal diseases, potentially influencing their decision to forgo medical consultation in the absence of severe symptoms.

Regarding treatment measures, 70.5% of participants reported using gastric mucosal repair drugs or probiotics, with examples provided such as "Compound Eosinophil-Lactobacillus Tablets" and "Live Combined Bifidobacterium, Lactobacillus, Enterococcus, and Bacillus cereus Tablets" (noting that these bacteria strains are probiotics). Additionally, 19.7% of participants utilized antibiotics (Figure 4). These data indicate a preference for pharmacological interventions, probiotics, and antibiotics as primary treatment modalities, likely due to their perceived effectiveness and convenience.

In assessing the effectiveness of treatments, 23.48% of patients noted symptom improvement at that time, however, the benefits were often not sustained, with instances of symptom recurrence observed. This finding highlights the limitations of current treatments and suggests a need for more enduring and effective interventions. Our product aims to address these shortcomings by providing a convenient and safe method to monitor symptoms proactively, thereby reducing the likelihood of recurrence.

When evaluating existing treatments, only 9.3% of participants deemed them "perfect", whereas over half (52.03%) expressed a preference for treatments capable of predicting and intervening in symptom development. Additionally, 39.02% of responders desired more persistent treatment effects with reduced recurrence rates, and 39.84% of them sought more convenient treatments, preferably requiring a single intake with a shorter treatment cycle. Furthermore, 20.73% indicated a preference for more affordable options (Figure 5). These insights inform our product's target by highlighting the demand for a probiotic platform designed to colonize the human gut, thereby mitigating dysbiosis-related diseases. We aim to develop minimally invasive, long-term solutions that continuously monitor the intestinal environment, intervene before symptom onset, and restore microbial balance proactively.

In the final two questions, participants were provided with a brief description of our product and subsequently asked about their acceptance and suggestions. Over half (54.07%) of the respondents found the product acceptable, while 39.43% chose not to evaluate it (Figure 6). This indicates a moderate level of public acceptance, though the prevalence of non-responses suggests that technical terminology may have impeded understanding. This underscores the necessity for ongoing product refinement and efforts to build social trust. Additionally, among the limited feedback received, some participants expressed concerns regarding the potential side effects of our products.

Overall, the questionnaire facilitated the identification of two key issues: intestinal diseases and gut microbiome imbalances are significant health concerns among Beijing residents, and there is a lack of long-term, predictive monitoring treatments in the market as the recurrence of symptoms further exacerbates patient inconvenience. Our products are designed to address these challenges. Based on the public's understanding of intestinal health issues, we will continue implementing educational campaigns.

We acknowledge the limitations of our questionnaire, including the restricted sample size and scope, and the potential for enhanced representativeness. We should minimize the use of technical language and give vivid examples to improve comprehension. Moreover, questions could be more comprehensive, addressing aspects such as the conditions under which participants experience intestinal discomfort, the frequency of symptoms, and whether stress exacerbates their intestinal issues. Moving forward, we plan to conduct expert interviews to enhance the professionalism of our project, ensure product safety, and establish greater maturity and responsibility towards the public.

In conclusion, despite the potential flaws mentioned above, this questionnaire activity has significantly contributed to honing our social science research skills and provided detailed insights that will facilitate the advancement of our products. Finally, we extend our sincere gratitude to the Beijing residents who participated in the survey, thereby supporting our research endeavors.

3. Interview with Gastroenterology Doctor Ms. Geng

From the literature we analyzed, we found that gut microbiota imbalance could lead to various diseases, most notably Crohn's disease and ulcerative colitis among Inflammatory Bowel Diseases (IBD). Focusing on gut dysbiosis, we're interested in its prevalence, especially in China in recent years. To ascertain the effectiveness of anti-inflammatory butyrate production in alleviating IBD symptoms, it’s crucial to know about the severity of these diseases from a health worker's perspective. In addition, we sought professional advice that compares the advantages and drawbacks of our probiotic to screening methods and other mature products in the industry such as antibiotics. This will enable our further development of the probiotic platform to fit practical needs through feasible means. With those queries in mind, we interviewed Ms. Geng, a gastroenterology doctor in a private hospital in China, United Family Healthcare (Figure 7).

From our interview, we learned the following key points:

1. The prevalence of intestinal disease has increased substantially since the last century, particularly in China. This could be attributed to people's deleterious dietary habits, particularly under the influence of fast-food culture, suggesting that apart from taking our "MetaboMender," patients also need to adjust their eating preferences to accommodate our treatment. Therefore, we could integrate lectures on nutritionally balanced diets with our project promotion during "Gut Alliance" online presentations and cross-talks with teams such as NMU-China and BIT-China. We could also design a tailored meal plan via mathematical modeling considering individual needs.

2. The intestinal flora is tightly interconnected with the immune system via signal transduction pathways as well as with our emotions via regulating neural transmitters such as GABA and serotonin through the gut-brain axis. Learning about IAA's potentially negative influence on the gut environment and its relatedness to Major Depressive Disorder supported our intention to regulate IAA's concentration when it's out of the optimum range.

3. We used to designate our probiotic product as a potential replacement for antibiotic treatment. Ms. Geng, however, stressed that doctors still resort to antibiotics for moderate and severe cases. Probiotics, on the other hand, are usually supplements rather than prescriptions unless dosages and strains are specified. The problem with current probiotic supplements is that individuals have unique gut microbiota, making the effectiveness of a standard set of probiotics vary between cases. Relatively limited varieties of probiotic strains result in limited effects. In addition, most probiotics fail to reach and colonize the intended section of the gastrointestinal tract due to the human body's natural barriers, as well as the competitive exclusion that protects our digestive system. Therefore, we slightly adjust our goal from treating people with sub-health gut environments to alleviating mild gut dysbiosis-related symptoms in people.

4. Interview with Synthetic Biology Researcher Mr. Zhang

Ms. Geng mentioned that synthetic biology has significant potential and future application value in the field of gastroenterology, especially in the area of de novo drug design. To gain a broader understanding of how synthetic biology is applied in various contexts, we interviewed Mr. Zhang, whose research focuses on the pharmaceutical potential of filamentous fungi through biosynthesis (Figure 8). This conversation offered valuable insights into how synthetic biology can be utilized to develop new therapeutic solutions, particularly in the biosynthesis of complex compounds for medical use. As a synthetic biology lab worker at the Institute of Medicinal Biotechnology in China, Mr. Zhang offered insights into improving the practicality of our project. After providing him with an overview of our background and experimental details, we interviewed him with two key focuses: general advice on synthetic biology experiments and specific feedback on our project design.

1. Project Feedback: Mr. Zhang praised the innovation and completeness of our project but suggested some technical improvements. For the IAA degradation part, he recommended using a single gene from the operon instead of the three-gene cluster (iadCDE), as this could improve expression efficiency and reduce plasmid burden. When we asked about adding three RBS sequences before the ORF to enhance expression, he agreed that this helps, but still emphasized the benefits of simplifying the operon to reduce the expression burden of the system.
2. Safety Switch: Mr. Zhang was particularly impressed with our safety switch design for its pioneering in China. He acknowledged that the safety protocols in local labs could lead to potential risks and appreciated our forward-thinking approach to containment and biosecurity, seeing it as a significant advancement for synthetic biology projects in the region.
3. Discrepancies in Experimental Protocols: We shared our concerns about encountering discrepancies between the experimental results and published literature. Mr. Zhang reassured us that this is a common occurrence in research. His advice was to remain flexible and conduct careful repetitions of our experiments, while critically integrating insights from multiple references to guide our work more effectively.

5. Collaboration

Being collaborative is essential for the iGEM community. BNDS-China provided fluorescent bacteria used for agar art for UCAS-China in appreciation of their warm accommodations, gifts, and bonds to BNDS-China since 2018. In addition, BNDS-China received the part PpChA for butyrate sensing from NMU-China and offered back a biosensor docking simulation for them.

Reference

Legan, T. B., Lavoie, B., & Mawe, G. M. (2022). Direct and indirect mechanisms by which the gut microbiota influence host serotonin systems. Neurogastroenterology & Motility, 34(10), e14346.