To optimize our team's project, we held an offline expert exchange meeting under the leadership of our PI, Principal Li Xun. During the meeting, we discussed whether thsS/R has specificity for thiosulfate and PphsA, as well as how to address the high off-target activity and large editing window of the CBE protein.

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Ultimately, after considering the teachers’ suggestions, we determined that thsS can only be phosphorylated in a thiosulfate environment, thereby activating its histidine kinase function, which leads to the phosphorylation and dimerization of thsR, giving it activity and allowing specific action on PphsA. Due to the inherently large size of the CBE protein, it exhibits off-target effects, low editing efficiency, and a large editing window, making standard CBE unsuitable for our project, which requires more precise and efficient editing. After reviewing a substantial amount of literature,we identified a small, efficient CBE protein called TadA-CDd, which was optimized from ABE through the PANCE (Phage-Assisted Non-Continuous Evolution) system, resulting in significantly lower off-target activity. We introduced the V106W mutation to further reduce its off-target rate, so we modified the original CBE gene in our system to the V106W mutant gene of TaDa-CDd.

To further refine our project and address the difficulties and uncertainties encountered during our literature review, we conducted a second online expert exchange meeting organized by our PI, Principal Li Xun.

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In this meeting, Teacher Li Yang explained the detailed signaling pathways of inflammatory factors and guided us in reading other high-impact literature to clarify our confusions. Teacher Li provided an in-depth analysis of the mechanism of action of the parasitic-derived AvCystatin (Av17), the drug protein selected by our team to serve as the treatment system in the overall system for IBD therapy. Parasites utilize highly effective immunomodulatory molecules to alter their host's immune system and reduce the inflammatory response against them. This mechanism is the basis for our choice of AvCystatin as the drug protein in the treatment system.

AvCystatin primarily targets macrophages, interfering with the host's immune system by regulating the expression level of IL-10, thus downregulating the inflammatory response against them. In the signaling events triggered by macrophages, AvCystatin is first absorbed by the macrophages, which subsequently induces the phosphorylation of mitogen-activated protein kinases (MAPK) ERK 1/2 and p38. The activation of these kinases can lead to the production of regulatory type 2 macrophages. The activation of ERK1/2 results in a rapid, transient phosphorylation of histone H3 at the IL-10 promoter, allowing it to bind to transcription factors such as Sp1 and STAT3, which are parallelly activated by p38. Notably, the expression of IL-10 in this induction is sensitive to tyrosine kinase and relies on the activation of both MAP kinases (which is in stark contrast to the expression of IL-12/23p40). Dual-specificity phosphatases (DUSP1 and DUSP2) regulate the induction of macrophages by AvCystatin, with DUSP1 specifically responsible for regulating the phosphorylation of ERK and p38, thereby controlling the expression of IL-10 in AvCystatin-regulated macrophages. In other words, AvCystatin induces and regulates macrophages via activation and inactivation pathways of MAP kinases. Additionally, AvCystatin can also induce the phosphorylation of transcription factors CREB and STAT3, regulated by phospho-ERK. Phosphoinositide-3-kinase (PI3K) can reduce the release of cytokines induced by AvCystatin, but downstream target AKT will not be activated after exposure to AvCystatin. We derived considerable benefit from this meeting and will continue to work hard to improve our project.

To understand the public's awareness of inflammatory bowel disease (IBD and analyze the social impact of education and outreach related to IBD, our team designed a simple survey questionnaire, which was widely distributed across various provinces in China. We hope to use the most research to improve our project. Additionally, our IHP group engaged in interviews with clinical and microbiological research institutions, interviewing senior colleagues. This time, we interviewed Dr. Jiang Lei, a surgeon from the First Clinical Medical College of Lanzhou University, and Ms. Mao Ting from the Gansu Provincial Institute of Biology.

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We collected a total of 500 questionnaires. Based on the data results, we reached the following conclusions: the public's understanding of inflammatory bowel disease is poor, with a generally low level of awareness, and the channels for promoting IBD need to be strengthened. Therefore, it is essential to enhance public education about IBD to improve awareness and understanding of the disease, thereby promoting health and preventive awareness among the public. For those who believe that awareness of IBD is unnecessary or manageable, more intuitive and easily understandable educational methods should be employed to help more people recognize the importance of IBD. Moreover, Dr. Jiang expressed his support for our use of engineered bacteria and synthetic biology to treat IBD. He is optimistic about molecular targeted research and hopes that emerging research findings can be applied to reduce severe cases, alleviate patient suffering, and avoid additional surgical trauma. From Ms. Mao, we gained valuable insights into the application of synthetic biology in the treatment of IBD. She also holds high hopes for molecular targeted therapy for IBD but emphasized the importance of biosafety.

In conjunction with the interviews from clinical and biological research institutions, our team held the third online expert discussion meeting. Due to the lengthy nature of our gene sequence, the probability of failure during gene insertion into plasmids is significantly high; thus, we mainly discussed how to address the oversized nature of our system. At the same time, we continued our interviews with clinical teachers, this time interviewing Dr. Ji Rui and Ms. Ren Qian from the First Clinical Medical College of Lanzhou University.

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Based on the teachers’ suggestions, we decided to split the plasmid, distributing the original large plasmid onto two high-copy plasmids with different resistance genes, each under 8000 bp, to reduce the difficulty of gene synthesis and electroporation, thereby ensuring the smooth progress of the experiment to some extent. During the interview, Dr. Ren provided detailed and valuable responses: whether the strains are colonized in the intestine, determining the concentration of anti-inflammatory factors released based on the severity of the patient's condition, and considerations regarding the accuracy of drug release locations, among others. Antibiotics and other medications can lead to many adverse reactions, and long-term use can reduce an individual's resistance and immunity. The tolerance level of individuals to medications also varies, making drugs unsuitable as a long-term solution. Additionally, there are significant challenges in the selection of microbial flora for fecal microbiota transplantation.

From our conversation with Dr. Ji Rui, we learned that most patients and their families are open to emerging treatments, and their acceptance of new therapies was an unexpected result prior to our interviews. As a result, we are more confident in optimizing our project and promoting our ideas and new treatment pathways to the public. We hope to ignite patients' confidence in recovery and provide them with encouragement and comfort.

In order to understand the TCM concept of inflammatory bowel disease, familiarize us with its prevention strategies.

Dr. Yao provide us with future improvement suggestions for our project from the perspective of TCM.Drawing on TCM therapeutic ideas has inspired us to delve deeper into researching gastrointestinal treatment targets, while also considering the possibility of looking further afield to explore the ultimate sites of inflammatory bowel disease.

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To gain a deeper understanding of the perspective of the integration of traditional Chinese and Western medicine on IBD, analyze the differentiation and treatment of UC and CD, and comprehensively understand the treatment approach of the integration of traditional Chinese and Western medicine for IBD, we interviewed Director Li Xuefeng of the Lanzhou University Traditional Chinese Medicine Culture Dissemination and Innovation Research Center to learn about his understanding of IBD.

Overall, TCM is an indispensable part of treating IBD. The future of integrative treatment for IBD is promising, but there is still a need for ongoing exploration and innovation in scientific research and clinical practice, continuously perfecting and optimizing treatment plans, with the fundamental starting point and ultimate goal always being to alleviate patients' suffering.

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In order to better focus on the physical and mental health of IBD patients, we interviewed Dr. Liu Jianbin, an experienced director of the mental health department, to explore how to build a solid mental health fortress for IBD.

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Looking ahead, we can enhance mental health education by using lectures, informational materials, online psychological support groups, and community outreach activities to raise awareness among patients and their families about the importance of psychological care. This can help create a caring and inclusive social environment, reduce discrimination and misunderstandings, and potentially offer personalized psychological intervention plans to provide multifaceted support and improve patients’ quality of life.

To broaden the team's knowledge of inflammatory bowel disease and better integrate the team's research into clinical practice, guided by the principle of responsibility towards IBD patients and the world, our team participated in the Gansu Province Inflammatory Bowel Disease Specialist Alliance Launch Meeting, the Gansu Province IBD Standardized Diagnosis and Treatment Guidelines Learning Session, and the 9th Gansu Province Intestinal Disease Forum and IBD Youth Forum, hosted by the Gastroenterology Department of the Second Affiliated Hospital of Lanzhou University on August 9-10, 2024.

This meeting primarily focused on expert lectures, classic case sharing, and case discussions, offering a rich and compact content experience and a strong learning atmosphere. We realized the importance of increased communication and learning. Promoting multidisciplinary collaboration is crucial for inflammatory bowel disease, and we hope to have the opportunity to participate in similar seminars in the future, gaining further insights into inflammatory bowel disease and contributing our strength to patients.

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In consideration of the deaf and hard of hearing community,Lanzhou University, in collaboration with Beijing Institute of Technology, Sichuan University, Jilin University, Hainan University, Nanjing University of Technology, and Zhejiang University of Technology, has launched an iGEM sign language science communication activity.

Through cooperation with universities in specific regions, the aim is to form a comprehensive network coverage model from east to west, south to north, and center, thereby expanding influence and promoting the enhancement of science popularization and innovation awareness.

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To raise awareness of inflammatory bowel disease (IB) among the elderly the northwest region, we chose Kang Le Hospital in Chengguan District, Lanzhou, as the venue for our outreach activities. This hospital serves a large number of elderly and caregivers, making it an ideal location to reach target audience. During these events, our team members provided detailed explanations of IBD to the elderly and their caregivers.

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We focused on explaining the symptoms, causes, and prevention methods of the disease, as well as the importance of early diagnosis and treatment. By increasing caregivers' awareness, we hope they can provide better care for elderly patients in their daily work. Our goal is to enhance societal awareness of IBD, help patients manage their chronic conditions more effectively, and ultimately improve their quality of life.

As a chronic intestinal disease, inflammatory bowel disease (IBD) has long course, and long-term medication can seriously affect the physical and mental health of patients In addition to general treatment, specialized care should not be overlooked. To understand the positive significance of nursing care related to IBD we interviewed Nurse Director Zhen Xiaoling from the Gastroenterology Department of Lanzhou University First Hospital.

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For IBD patients, effective nursing measures include psychological support, lifestyle management, medication management, and pain management. Providing appropriate and quality care is significant for patients with inflammatory bowel disease. To strengthen research collaboration within the gastrointestinal research team and to mutually learn from the new ideas and concepts of different teams, we initiated the Gut Alliance in collaboration with Tsinghua University.

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The Gut Alliance aims to:
1.Promote Academia:Enhance the depth and breadth of research on intestinal diseases and accelerate scientific discoveries.
2.Support the Medical Community:Provide clinical doctors with the latestresearch findings to optimize treatment plans and improve patient outcomes.
3.Influence Society:Raise public awareness of gut health, advocate for healthy lifestyles, and reduce the incidence of intestinal diseases.
To facilitate in-depth communication and sharing of expertise and the latest research findings in the field of synthetic biology, and to promote academic integration and collaboration between our two universities, we created content in a question-and-answer format based on synthetic biology and our respective project themes. We hope that this engaging and interesting format will stimulate a strong interest in synthetic biology and promote its development in practical applications

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This collaborative recording activity has built a bridge for academic communication between the universities, allowing both sides to fully integrate their advantageous resources and form a powerful alliance to explore the unknown mysteries in the field of synthetic biology together. The question-and-answer format of the podcast content makes it easier to convey specialized knowledge clearly and accurately, enabling listeners to better understand complex scientific concepts and making a positive contribution to the popularization of science and scientific education.

The team has conducted solid theoretical research on inflammatory bowel disease (IBD) and has gained a comprehensive understanding of its pathogenesis and prevention strategies. As a result, we created an IBD knowledge manual to out public science education in communities, helping the public to correctly understand IBD.

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Our work has not only increased public awareness of IBD but also highlighted the importance of early diagnosis and timely treatment. By dispelling public misconceptions and biases about IBD, we have promoted societal support and understanding for patients. In addition, we encourage the public to adopt healthy lifestyles to facilitate early prevention of IBD. These efforts will contribute to reducing the overall prevalence of IBD in society and alleviating the medical burden on the community.

In order to popularize synthetic biology and IBD, we have successively conducted cloud teaching, Western Sky Association teaching, Teaching Association teaching, Lanzhou Baishuxiang teaching, Luotuoxiang teaching, Linjiazhuang teaching, Yuzhong County Picture Book Museum teaching, Zhangye City teaching,Longnan City teaching, Lanzhou University Affiliated Primary School teaching, etc.

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To gain a deeper understanding of advancements in intestinal technology, companies in the field of IBD research, and how they leverage technology to drive the development of innovative therapies, we collected valuable information regarding research findings, technology applications, and future directions through Professor Lu's expertise. Additionally, we drafted a preliminary business plan in conjunction with our project.

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During the series of activities and interviews with numerous stakeholders, the team members will promptly summarize the meaningful content (in both Chinese and English) and then publish it on various online platforms in the form of tweets or videos for public education. As of September 25, 2024, the total viewership across all platforms has exceeded 10,000.This season, the team has released the following content:24 posts on the WeChat public account platform9 videos on platforms such as Douyin, Bilibili, and Xiaohongshu, including 6 popular science experiment videos, 2 interview videos, and 1 sign language video.

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