Integrated

      For the purpose of applying the results of human practice to experimental design, feeding the results of experiments back to human practice, we have conducted plenty of interesting and innovative activities. In addition to questionnaire collection, large-scale popular science outreach in high schools and kindergartens, we attended joint conference of universities and participated in national academic conferences, etc. We also innovatively focused on the practice of local people solving local problems. Therefore, the Longan township, Fuqing, which fits our mission was chosen and a complete solution to Fuqing’s problem through experiments, biostatistics, modeling, deep and extensive visits was achieved. It is worth mentioning that we pay attention to the application of the results to the industrial transformation as well. Our team members have completed the detailed and scientific business planning and initial production of products, participating in various entrepreneurial competitions to improve the plan.

Questionnaire Survey

1.Questionnaire Survey
Fig. 1.1 The result of Questionnaire Survey    To implement our experiment more effectively and thoroughly, we collected 102 questionnaires and analyzed the consequences as follows:
      About the experience of mental or physical health problems, anxiety took the largest proportion with 65.69% according to the records, followed by mood swings and insomnia with 51.96%, 50.98% respectively. For those who have these problems, above 62 percent group of people tend to sport or adjusting their own dietary habit (46.08%), but seldom rely on drugs. Hence, with only 14.71% of them have taken medicine to fight against sleeplessness. To investigate whether the existing group who have the metal problems, we asked them how they like the current products to help them. According to our discovery, people mainly reckon those pills or drugs could basically satisfy their demand.
      Apart from that, some have been interested in products related to GABA, like GABA drinks and candies. Simultaneously, Longan is getting its focus as a significant tropical fruit with high nutrient value. As more than half of our subject have never heard of GABA, we find it important to tell more about its functions of improving sleep quality. Since an overwhelming number of individuals believe that GABA is potential after we introduce it to them, we determine to spread the knowledge about GABA and our project that contributes to addressing metal problems via this chemical substance with gene editing and systhetic biology.
      More than half of the people have heard of synthetic biology according to the consequences of questionnaires, and most of them learned about it through the Internet and people around them. Therefore, we can conclude that synthetic biology has attracted increasing attention as it is an international frontier and has a wide range of prospects, and people are most interested in the theory that synthetic biology can be used to synthesize the materials they want for living things. Although 95.1% of people have heard of gene editing technology, people do not know much about gene editing technology in synthetic biology, and many people know about transgenic technology but do not know the difference between it and gene editing. 53.92% of respondents were critical of gene editing as a technology that has both positive and possible negative effects on human health. So for the public, they are more interested in the safety of gene editing products and secondly in the efficiency of technology.
      Summarizing all the results we collected, we determine to devote to cope with insomnia by means of GABA gene editing and systhetic biology to help individuals get back into their normal lives. Furthermore, our team will work to develop the functions of GABA and spread the knowledge to more so that they could benefit from our project and product, both with scientific and medical value. Last but not least, by virtue of the iGEM platform, we will tell people of all the ages about the charm of systhetic biology and its related researches.

Expert Interview

2. Expert Interview
①2022.07.07 Professor Longtao Zhang
Fig 2.1 The team members visited Professor Zhang Longtao       At 11:00 a.m. on July 7th, we visited Professor Zhang to consult on the establishment of project proposition and the improvement of experimental plans. Professor Zhang is committed to the organic combination of teaching, scientific research and industrial needs, who has maintained frequent cooperation with multiple large and medium-sized food enterprises and provided valuable suggestions for the development of our project.
      First, Professor Zhang pointed out that we may as well consider whether the project is viable from the perspective of investment. This requires taking the current feasibility of Longan cell production into account. Additionally, we need to compare and analyze the microbial production (e.coli, lactic acid bacteria, etc.) on the current market to understand the advantages of Longan in terms of efficiency and costs. Finally, we need to decide whether the production is extracellular or intracellular.
      As for the market, Professor Zhang told us to consider the market positioning, prospects, target users, and the specific form of the product. Furthermore, it should also be clear how much GABA products should be consumed when they functions, and how to promote GABA products to promote sales.

②2022.7.15-7.17 The most beautiful expert who works locally in Fujian province--- Weiping Zheng Fig 2.2 The picture shows Expert Weiping Zheng having discussions and exchanges with other members       Zheng has rich experience in exploring new types of agriculture, who established the Longan brand of Yuxi as well as Yuxi Longan production base. He built a green ecological farm integrating large-scale, commercialization and ecology, and opened up a path of poverty alleviation through scientific and technological enrichment and green development. Zheng has won the honors of "2021 Fujian Most Beautiful Local Expert" and "Rural Revitalization Talent". Particularly, he pays attention to the improvement of Longan quality, insisting on the production of green products, honest management and firmly achieves NO MEDICINE in orchard and NO DAMAGE during logistics to form a good reputation for its brand, which constantly raises farmers' income and pave the way for the journey of rural revitalization.
      Weiping Zheng's successful case provides new ideas for our unique local solutions, and also reminds us to establish our own brand "High GABA Longan" in the subsequent product processing. He told to our team eagerly, "the revitalization of the agriculture development needs the power of youths. Remember, never stop learning, or we will be backward. And don't be afraid to go out and give it a try". The youth should have the courage to blaze new trails in boosting rural revitalization of youthful vitality.

③2022.7.15-7.17 Fuqing Agricultural Technology Association Secretary General Yu Zuxing Fig 2.3 Zuxing Yu, secretary general of Fuqing Agricultural Technology Association, is having a discussion with team members       We consulted Mr. Yu about the current state of the Yuxi Longan industry and the authorities' attitude toward integrating gene-editing technology with agriculture. What he told us was as follows.
(1) For the problems of primitive agriculture
① Limit chemical fertilizers and pesticides, and now encourage organic fertilizers
② Control the Longan shoot problem

(2) The causes of the original agricultural problems
① The influence of historical years
      Mainly caused by improper management, production exceeded the standard and other reasons caused by the second year of a significant decline in output. If more fruits are produced in the first year, or if the proportion of leaves and fruit is not correct, the quality would be disappointing. Apart from that, it will also affect the output of the second year, which is the formation of "big year" phenomenon.

② Winter shoot flushing phenomenon
      In order to solve this problem, local farmers have to hedge the shoot phenomenon to control, mainly through ethephon (30ML ethylene with 100 kg of water to spray longan) to accomplish. At present, the local technology has been developed and mature, which has a good effect. Farmers controlled the winter shoots so that sugar could accumulate without affecting the next year's production.
      Mr. Yu pointed out that variety is the foundation of the fruit quality. Yuxi Longan has a long history, benefited from unique geographical conditions and varieties. In order to develop Yuxi Longan to a new level, it is necessary to attach importance to science and technology, and pay attention to the reform and innovation brought by Longan Science and Technology Academy. He strongly supported our project and expected our technology to drive the growth of local Longan industry.

④ Doctor Zhenshan Lin
Fig 2.3 With Doctor Zhenshan Lin Interviewer: Dr. Lin, we now know the ingredients for making GABA drinks, but how much each ingredient do we really need?

Lin: It depends on your own needs. For example, how many bottles of drinks do you want to make?

Interviewer: About 50 bottles.

Lin: You can actually calculate the amount of each ingredient. The amount of essence and other additives are usually a few parts per thousand. You can figure out how much each ingredient to buy according to the volume of each bottle of drink.

Interviewer: Got that. But can we know the specific number for the thousandths and ten-thousandths you just mentioned?

Lin: It doesn't need to be that clear and the data can't be determined now. After you buy the raw materials, I will taste the drink consists of them with different proportions and then I can decide the content and its concentration.

Interviewer: OK, we will first purchase the raw material in a certain quantity. Which specifications do we need to purchase the centrifuge tube? What is the quantity?

Lin: There are a lot of centrifuge tubes in the laboratory. You can buy some new ones and the specifications should not be too small, since we will use centrifuge tubes to test the drinks later and a slightly larger capacity is of requirement.

Interviewer: What type of pectin would be a good drink for us to use?

Lin: It should be oligomeric, which mainly plays the role of stability and condensation.

Interviewer: You told us that Guangzhou Baizhong Flavor should be used for beverage flavor, but we didn't find pectin of this brand on the shopping platform. Which pectin should we use instead?

Lin: Guangzhou Baizhong Flavor is what I used to do a flavor of the drink. I also used a MANSI flavor to create the drink flavor that is also sound, you can look for it.

Interviewer: Yet, unfortunately we did not find any related merchants selling MANSI flavor.

Lin: That doesn't matter. As far as I know, the focus of your project is to extract GABA from Longan and apply it to other aspects, so the taste of your drinks does not need to be very strong. Even if you don't add essence, there is no problem.

Interviewer: OK, thank you for answering our questions, Dr. Lin.

⑤ Head of Herbal Tea Group, entrepreneur Chunsong Lin
Fig 2.5.1 The picture shows the FAFU-China team(first left) communicating with the boss of Herbal Tea Group (second left).       First of all, Mr. Chunsong Lin invite us to taste Fuding white tea and taught us how to open our own taste experience before and after the taste. Then he led us to taste the herbal tea newly developed and made a simple comparison with the Longan drink sent by our team. Herbs, herbal tea sweet not always stay in the mouth, let a person want to drink again after a small bite. As with sugar and fructose, the sweetness of Longan drinks cannot remain in mouth, then we would like to try other sugars that sucrose can be replaced, also regarding to change the processing technology. At present, the stores under Herbal Tea company join Eleme and Meituan takeout so as to ensure the stores have their own industrial chain to supply food materials. Mr. Lin talked about the key elements of entrepreneurship is not only to keep pace with the times, but also to make our products in line with the public taste, to achieve the ultimate pursuit of taste.
      The main content of the drink is GABA, a health care beverage for insomnia. At present, the GABA content of the beverage containing Longan is not our goal. Can we try to add GABA powder as substitute to check the taste and other factors? The addition of longan is 25% and 30%, but the fruit flavor is not enough (additive - health). After people taste it, they commended that the flavor of sour and sweet are not harmonious and the astringency is heavy, which recommends us to adjust the taste.
      Our drinks face the same problems as our company: the effective ingredients change when the temperature is high, and the taste is affected. May consider the development of freeze-dried powder or other.
      At present, the beverage emphasizes 0 sugar, 0 fat and 0 calories. If you drink a cup of Longan GABA drink before going to bed, whether you think that drinking too much sweet before going to bed is not healthy.
      Finally, Lin invited the FAFU-China team to work together in the future to participate in the company's new beverage tasting. Fig 2.5.2 The picture shows the successful end of the exchange. Second from right: Luo Xun, an outstanding graduate of Shanghai Jiao Tong University, who enters the beverage company later. ⑥ Dr. Chen Chao, Inpatient Department, Fujian Provincial Hospital
      On the afternoon of July 28th, 2022, team members of FAFU-China went to the Inpatient Department of Fujian Provincial Hospital to interview Dr. Chao on the treatment of anxiety disorders, sleeplessness and depression.
      During the conversation, we learned that at present, the hospital received less depression patients but more with insomnia. The main causes of insomnia are: difficulty falling asleep, waking up early, sleeping nightmares, snoring, among which anxiety patients are mainly middle-aged women experiencing menopause. In the wake of the pandemic, there is a modest increase in the number of patients coming to us for treatment of these conditions.
      At present, the mainstream treatment methods are divided into two kinds, one is through the psychological counseling of the patient's language, the other is the more effective method is through drug treatment. At present, new sedative drugs (Xnox, dexzopicron tablets) are mainly used for drug treatment, and some more traditional antipsychotic drugs (Alprazolam, Estazolam) are also applied. As one of the main inhibitory neurotransmitters in the central nervous system, GABA aminobutyric acid can affect the central nervous system via neurotransmitters such as GABA to achieve the role of treating anxiety disorders and insomnia disorders. The dominant side effects, however, are: dizziness on the second day, want to sleep on the second day, long-term use is easy to addiction, high recurrence rate after treatment (solution: adding antidepressants in the later stage). Yiming Ma, our team member, who talked about her symptoms of anxiety during the college entrance examination after taking drugs to improve is not mitigated, asked whether the drug could achieve the therapeutic effect owing to the psychological effect in the treatment of no practical utility. Doctor Chao explained that placebo, which we think, is not used in the real treatment process. Most of the treatments for anxiety and depression are really pharmacological effects. There is a lot of crossover in the presence of drug therapy in depression, anxiety, insomnia and other symptoms. Fig. 2.6.1 The picture shows the team members talking with Dr. Chao Fig 2.6.2 At the end of the interview, the team members took a photo with Dr. Chen Chao