Team:XJTU-CHINA/Human Practices
Integrated Human Practices
Our human practices are quite active through the whole project.
In the inspiration stage of our project, an initial data collection about the people's sleep quality and the experts interviews, indicate the high rate of insomnia with limited solutions, which inspired us to improve people's sleep quality and mental state as much as we can. After market investigation on sleep-aids therapy and discussion with industrial producers of fragrance products, we launched our project that selective microbial production of two fragrance molecules.
In the implementation stage, we encountered many problems especially product design and biosafety concerns since our product needs to be used at home in the daily life, which cannot be effectively solved by our own knowledge. Therefore, we consulted several relevant experts, other iGEM teams nationwide etc. and applied their suggestions into our product design, to avoid bacterial leakage, undesirable odor problem as well as App design.
We also planned for the future. And we got a lot of inspirations about smart sleep monitoring and entrepreneurship, trying to further promote our project in the future.
Inspiration Stage of Project
Sleep-related Questionnaires
A questionnaire was launched in order to learn the sleep quality of people and their attitudes towards our product. The respondents of our survey are mainly students and middle-aged workers and the total number is 1315.
Results of the questionnaire on sleep status:
- The average sleep time of the population is 6-8 hours. But about 75% of them suffer from sleep problems, including difficulty to fall asleep, insomnia, and shallow sleep. In addition, more than half feel sleepy when they wake up in the morning.
- It is widely believed that the main factor affecting sleep is emotion and stress. A variety of methods are tried to improve their sleep quality, including exercise, diet, music etc.
- But for scent products, such as aromatherapy, only nearly half of people think they can effectively improve sleep quality and mental state, while about 47% of the respondents would like to try aromatherapy products to improve sleep, but the acceptable price is expected to be below 300 RMB (~40 USD).
Interviews with sleep-related experts and market investigation on therapies
Interview with Prof. Jiangang Long, an expert in sleep disorder
We found that sleep disorder is quite common nowadays after investigation. We want to learn more about its possible impact, so we referred to Prof. Jiangang Long for help, who is an expert of this field and also the Vice Dean of School of Life Science and Technology in XJTU.
Prof Long mainly engaged in the anti-fatigue effect of natural extracts on sleep disorders and their mitochondrial mechanisms. He told us that the lack of sleep can cause rhythm disorder, oxidative damage, decreased brain function, Alzheimer's disease and so on. Some resent studies have found that telomerase activity, that is highly correlated with longevity, may also be affected after sleep deprivation treatment. In his opinion, sleep loss is normally reversible, but this revision cannot be achieved in the extreme cases, so Prof Long suggested to get more sleep for recovery, but if the situation is too severe, you need to change your habit and take some medicine.
After talking with Prof. Long, we learned more about the possible harm of insufficient sleep from an experimental point of view, and realized the importance of adequate sleep for health, which inspired us to consider how to solve this problem.
Interview with Prof. Wei Dang, an expert in therapy of sleep and mental disorders
How to solve the problem of insomnia? We consulted Prof. Dang, the Director of Xi'an Mental Health Research Center. He gave us his own views on the treatment of sleep disorder. He believes that our project is of great value in application.
He said that in recent years, many people suffer from sub-health such as fatigue, and irritability, which will affect their sleep quality. The first choice is drug, but long-term intake of drugs may cause harm to the body, and even resulting in drug-dependence. However, some other assist therapies of insomnia, such as medicine pillows, traditional Chinese medicines and TMS TCD static intracranial stimulation is less harmful but effects are not obvious.
After the conversation with Prof. Dang, we realized the necessity of research on assist therapy of insomnia, and we investigated the existing sleep-aid methods in the market, which is listed as follows:
Sleeping Aids
| Aromatherapy | Medicine (e.g. methylalanine tablets, chlormethazone tablets, etc.) | Health products (e.g. melatonin, etc.) | Intelligent devices (e.g. smart watches, smart mattresses) | Physical sleep aids (e.g. eye patches, sound-proof earplugs, etc.) | |
|---|---|---|---|---|---|
| Production costs | ★★★★★ | ★★★★ | ★★★★ | ★★★ | ★★★★★ |
| Raw material availability | ★★★★★ | ★★★★★ | ★★★★★ | ★★★★★ | ★★★★★ |
| Purchase cost | ★★★★★ | ★★★ | ★★★ | ★★ | ★★★★★ |
| Effect | ★★★★ | ★★★★★ | ★★★★ | ★★★ | ★★★ |
| Side effect | ★★★★★ | ★★ | ★★★★ | ★★★★★ | ★★★★★ |
Nowadays, there are several kinds of sleep-assisting products in the market including physical and chemical way. Patch, foam ear plugs, night light, intelligent mattress, intelligent band and special pillows is physical aids while aromatherapy, enzymes, health-care products and melatonin are chemical aids.
From the manufacturer's point of view, the production costs of physical aid and aromatherapy are lower compared to intelligent machine and medicine. And as a consumer, he prefers the product with effective result and reasonable price, that is, the product with high performance cost ratio. Some physical aids such as patch and foam ear plugs are cheap and have positive effect to some extent, but they are not enough for some situations. Aromatherapy is an appropriate choice with affordable cost and effective result. What's more, aromatherapy will not cause side effects compared with some drugs. From the data reported by CBNDATA, most of the people prefer special perfume to promote sleep.
In all, taking all factors into consideration, aromatherapy are one of the best choices for assist therapy of sleep.
Then, we started to consider what kind of fragrance to produce, so we conducted further market research.
Visits of Fragrance-producing companies
Xi'an Natural Field Bio-technique Co., Ltd
Based on the above preparation, we plan to alleviate insomnia by producing fragrance. As we know Xi'an Natural Field Bio-technique Co., Ltd is rich experienced in the production of plant fragrance, so we visited them and discussed our project with Manager Yang of Quality Department and learned a lot about how to improve our project.
First, Manager Yang thought highly of our bacterial fragrance generator project. He told us currently the most conventional method for fragrance extract is mainly physical process with easy operation but high cost and low yield. And if microorganisms are used for fragrance production, safety is the most important consideration. Second, as a businessman, Manager Yang kindly advised that we'd better to think more about our cost and whether people can accept our product or not. Furthermore, Manager Yang suggested that we should make sure the efficacy of linalool and limonene in solving sleep-related problems. In conclusion, if we want to put our idea into production, the safety, cost of our product and the acceptance of customers must be taken into consideration.
Thanks to Xi'an Natural Field Bio-technique Co., Ltd and Manager Yang, we realized our project still needs improving. And we got an idea about what we should do in the next step of our human practice.
Thanks to Xi'an Natural Field Bio-technique Co., Ltd and Manager Yang, we realized our project still needs improvement. Based on his suggestion, a questionnaire were conducted to investigate the attitude of customers on the safety, cost of our product in the following human practices.
Key points of discussion with quality department Manager Yang:
- The process of the herb extracts is mainly physical process with high cost. In view of the advantages of high production efficiency and low cost in bio-production, we believe that the use of biosynthesis to produce fragrance can optimize the existing plant extraction methods to a certain extent.
- If we use microorganism to produce herb extracts, we should take safety into account. We should also think about our cost and how people accept our products. We realize that it is necessary to investigate the public's acceptance of microbial products. Based on this, we conducted a questionnaire survey on this question.
Results of the questionnaire on microbial products:
For microbial products, most people agree that they are willing to try and use home systems that are based on microbial systems, but to ensure product safety. In addition, people's awareness of microbial safety issues is still lacking.
- Manager Yang suggested that we should make sure the efficacy of linalool and limonene in solving sleep-related problems
After discussing with Manager Yang, we consulted the relevant literature and confirmed that linalool is reported to promote sound sleep with peaceful mind, and limonene allows people to feel less sleepy and distraction in the morning, indicating limonene and linalool can work effectively on improving people's mental state and sleeping.
Xi'an Yuensun Biological Technology Co., Ltd
"Microbial synthesis will be an admirable way to reduce production cost and enhance productivity."--- Mr. Jin, General manager and executive director
Xi'an Yuensun Biological Technology Co., Ltd. with a famous brand of "YESHERB", is a leading plant extracting herbal formula supplier in China. They devote to providing not only Chinese herb extract but efficient way of use. Their products range from Plant Extracts, Mushroom Extract, Fruit Powder, Vegetable Powder, Super Foods.
Xian Yuensun Biological Technology Co., Ltd. with a famous brand of "YESHERB", is a leading plant extracting herbal formula supplier in China. They devote to providing not only Chinese herb extract but also recommending effective ways to use these herbs. Their products range from Plant Extracts, Mushroom Extract, Fruit Powder, Vegetable Powder, Super Foods.
The General Manager and Executive Director of Yuensun Biological Technology Co., Ltd Mr. Jin, thought highly that our microbial production technology for fragrance will reduce the cost and enhance productivity, because the production cost and productivity of plant fragrance are largely depend on the accessibility and amount of raw material, while our microbial method will solve the problem. Nevertheless, our project still has problems that needs to be solved. Firstly, the safety of our engineering bacteria deserves more attention. Furthermore, we should focus on the cost reduction in later development stage of the project, as only low cost can bring more benefits to the society.
Key points:
- During the production of plant extract, the production cost and productivity severely depend on the accessibility and amount of raw material. It may be possible to solve this problem through biological production.
- If we only use microbial metabolites instead of making microorganism as product and sending them to the society, microbial contamination can be avoided. In response to this problem, we consulted Jiping Zhao, who believed that it's very complicated to implement cell free. At present, we can ignore it, instead, we can ensure biosafety from strict sterilization and hardware design. Of course, cell free can be one of the future directions of our project.
- The market condition of our product from the engineering bacteria deserves more attention. Having looked at the table that is previously summarized, he expressed his appreciation and suggested that we should continue to collect the relevant market data.
- In later development stage, only minimize production cost can this project bring more benefits to the society. At present, we believe that the cost of culture medium can be reduced by acclimatization of strains to grow normally in low nutrient medium.
Implementation Stage of Project
MEETUP
In the process of the project, we also encountered some problems. We have learned a lot by communication with other teams. They also offered some great suggestions to improve our project.
Southwest Meetup
- Time: 2019.5.1-2019.5.3
- Location: Sichuan, China
- Participates: iGEMers of Xi'an Jiaotong University, Sichuan University, University of Electronic Science and Technology of China, Third Military Medical University, and Sichuan Agricultural University, etc.
Our team participated in Southwest Meetup of five iGEM teams in Southwest of China on May 1. We made presentations to share our ideas with each other. At that time, we didn't launched our project. iGEMers from Sichuan university and University of Electronic Science and Technology helped us a lot in terms of project feasibility. Meanwhile, Ambassador Zhang taught us how to make a fabulous human practices and highlighted the importance of integrated work. And she also pointed out the biosafety issue of our project which we ignored of at the beginning. This has had a huge impact on the design of our hardware. We have thought of a variety of methods such as setting up separation layers to solve the problem of bacterial leakage. We have benefited a lot from this meeting.
The solutions we envisage for biosafety:
- Insert isolation chamber in hardware to separate activated bacteria from inactivated ones.
- Blanch bacteria by adding bacteriostatic agents after use.
- Use nutritionally deficient E. coli to reduce its activity and to ensure biosafety
- Use suicide mechanism in E. coli to ensure biosafety.
Shaanxi Meetup
- Time: 2019.7.19
- Location: Xi'an Jiaotong University
- Participates: iGEMers of Xi'an Jiaotong University, Yan'an University, Northwest University etc.
XJTU-iGEM team held a meeting with the YAU-iGEM team and Northwest University 2019 iGEM team in Xi'an Jiaotong University.
Each team gave a brief introduction of their projects, followed by in-depth discussion and communication. Our team members put up with some suggestions to improve the project of YAU-iGEM team. Most importantly, we decided to build a Northwest database of frequently-used genes or parts among iGEM teams in Northwest China, to facilitate the future teams. This is five-year plan, and we have synthetized and build a DNA database with 150kb DNA lengths.
The members of each team not only deepened their friendship, but also started several cooperation with each other after this meetup.
Conference of China iGEMer Community
- Time: 2019.8.19-2019.8.23
- Location: Shenzhen, China
- Participates: iGEM China community
We participated in China community meetup organized by Shenzhen Institutes of Advanced Technology. We have received many useful suggestions from other teams and experts in the field of Synthetic biology.
iGEMers from HAZU thought that we needed to pay attention to the continuous culture of bacteria. After this meeting, we interviewed Professor Zhang (major in microbiology) to solve this problem an idea to make a "dry package" come to our mind. In this event, we found the undesirable odor of E. coli is also a big challenge. Students from Northwest University suggested us use Crisper-cas9 to block related genes and judges encouraged us to do cell-free system. These suggestions inspired us to think further and wisely. In addition, we also adjusted the direction of modeling to conduct the stability analysis of the ligh-control switch circuit according to the advice of the judge.
Besides our project, we received advice on our presentation and posters. We learned a lot of typography and beautification skills at the meeting. We really appreciated everyone's help and advice and our project benefited a lot form this meetup.
Interviews
Circuit Design
After we discussed with Prof. Hua Dong Liu, an expert in synthetic biology, about our ideas and circuit design of the project, he suggested that our circuit could be further simplified shown as followings. And this simplification greatly reduced the complexity of our experiment, which is very meaningful and helpful to our project.
After listening to the teacher's suggestion, we think that this simplification can greatly reduce the complexity of our experiment, which is very meaningful.
Hardware and Biosafety
In implementation of project, we found that the undesirable odor produced by E. coli metabolism would affect the smell of our product. Besides, our product need to meet the bio-safety standards for family use. And we have no idea about how to solve the problems, then we referred to Prof. Yali Zhang, an expert in microbial culture.
She provided us with some feasible suggestions on the hardware and product design, and we applied her advices into our design to improve our project. This consultation helps a lot to our follow-up process and its future development.
Key points:
-
Replacing the organic nitrogen source in the culture medium will reduce the undesirable odor caused by E. coli metabolism.
Based on this recommendation, we replaced LB medium in the laboratory with YT medium with high C / N ratio. We cultured E. coli overnight both in LB and YT medium.
Finally, when the difference of OD600 between the two bacteria liquid is only 0.07, we take the supernatant after centrifugation, and measure its pH value.
The results are as follows:
It can be seen from the results that when the concentration of E. coli is basically the same, the amount of ammonia produced by E. coli cultured in nitrogen reducing medium is less. Therefore, it can be concluded that nitrogen reducing medium has a certain effect on solving the odor produced by E. coli.Figure1. PH value of medium（before culture）Figure2. PH value of medium （after culture）
- In order to make the product more convenient to use, E. coli and culture medium can be made into dry powder.
- In order to ensure biosafety, ultraviolet radiation is not enough for sterilization. High temperature sterilization is the best method, if it cannot be achieved, chemical reagents like some blanches can also be considered.
- After knowing the purpose of our project and the existing problems in product detection, Prof. Zhang suggested that we use low-temperature culture to achieve product enrichment. Based on this, we cultured E. coli at 28 ℃ and finally synthesized linalool synthetase LGF.
- The inorganic ions in ordinary medium can basically support the growth of microorganisms without additional supplementation.
- In addition, Prof. Zhang also raised a question about our experimental design: Whether the bio-heat produced by E. coli metabolism will cause the temperature-controlling switch to fail to work needs further research? So we conducted the experiment to verify this issue and found the biological heat produced by E. coli in our system cannot reach 42℃ at all, so it is supposed that our switch system can work efficiently. Besides, we still add a temperature detector in the hardware design to ensure the system work properly.
Questionnaire on APP
In order to make our products more convenient to use and ensure its biosafety, we decided to use APP to achieve remote control. To better understand the future market demand and acceptance of our devices and APP, we designed a questionnaire and obtained the following conclusions based on the survey results.
- Currently, about half of people have used smart control appliances. And about 65% of people think that remote intelligent control appliances can bring more convenience to life.
- For the smart-home coupled APP, people pay more attention to the characteristics, which includes the home appliance control, intuitive and easy to understand and simple and functional of the interface. In addition, people hope that APP has the following functions: remote control of smell release and stop, timely reminding of the replacement of related accessories (bacterial package) and promptly reminding the product of the current security risks.
- For the product, the user wants the replacement frequency of the product accessories (bacteria package) to be as low as possible, such as once a month or even once half years.
- About 75% of people are willing to purchase our products and download related Apps. Besides, the price that most people can afford is between 50 and 200 RMB (~7 and 28 USD).
According to the results of the survey, our products and coupled APP have a certain degree of market acceptance, which laid the foundation for our future market. In addition, the respondents provided several suggestions for the improvement and optimization of the function of APP in the future.
Plan for the future
Sleep monitoring
After most of our projects have been completed, we found that there was no reliable way to demonstrate that our products did play a role in improving people's mental state or alleviating insomnia. Therefore, we plan to design a device that can effectively detect the quality of sleep in the future, which can also couple with our APP so that the users will know their sleep quality.
However, we are not experienced in this field, so we consulted Prof. Jin Xu, a specialist in biomedical engineering and sleep monitoring.
Key-points:
- At present, most of the Apps related to sleep used in the market have the function of sleep monitoring. However, most products rely on the body motion as an indicator, which is lack of accuracy.
- For clinic, sleep monitoring is more scientific and accurate, and the principle of clinical testing and household products are the same, so electrocardiogram (ECG), body position and electroencephalography (EEG) method used in clinic can be used to monitor sleep quality in our future project.
- Prof Xu suggested that ECG is easy to achieve, but it is rough analysis. EEG detection can basically achieve accurate analysis, but it is uncomfortable to use. Taking all together, we plan to combine EEG, ECG and body position analysis to develop our project.
- Mental psychology, nerves, and breathing can cause sleep disorders. At present, many hospitals pay more attention to this issue and begin to set up sleep-related department, which is the future direction.
Sleep disorders and their detection require more attentions. How to achieve effective sleep monitoring is challenging. Thanks to Prof. Xu, we find a primary solution for sleep monitoring in our future project.
Entrepreneurship
Based on our previous market research, we believe that our product has a great potential to enter the market or apply for a patent, so we consulted an expert Dr. Lv in transformation of scientific results into practice. Mr. Lv believed that our project is novel and has high application value, as smart home becomes a hot topic currently. Our project achieved the selective fragrance production to improve sleep quality coupled with APP remote control, which is quite innovative. However, if the product is about to enter the market, the following points should be considered:
- Improve existing production efficiency and reduce the costs
Product commercialization needs mass production. At this stage, the product can only achieve a small amount of yield in the laboratory. If the conditions change, a functional production system may not able to work. Therefore, a lot of tests are needed before the scale-up and mass production. - Needs an attractive product positioning
This project can be considered as an air freshener or sleep facility, but it is more recommended as a smart home to promote, which is more attractive.
Others
Apply for the independent laboratory for iGEM
At present, most iGEM teams do not have independent laboratories for their project. They basically carry out iGEM projects in the laboratories of professors. At first, we did the same. However, in the process of our project, many junior students contacted us and told us they are willing to join iGEM next year. We began to realize that implementation of project in the shared space with other non-iGEMers is less efficient and is not conducive to the inheritance, so we have applied for an independent laboratory space for iGEMers from school, which is located in the Room 104 of our department, and equipped with sufficient instrument for our experiment. Up to now, many junior students have come to visit our laboratory. iGEMers also showed the experimental operation of synthetic biology to them. The junior students said that they can try their own ideas boldly in such a free environment and feel the joy of scientific research.
Berkeley iGEM Communication
During the summer vacation of 2019, some members of our team participated in the Summer Exchange Program at UCB, where we had the opportunity to meet Prof. Terry Johnson, a professor of bioengineering who was previously a judge of iGEM. Firstly, in his lecture, he shared with us the iGEM project Berkeley had done. He especially emphasized how ideas came into being, which made us realize the importance of learning from life. In addition, he also explained the problems that should be paid attention to in the process of product design from the point of view of engineering design, which is very helpful to the construction of our hardware.
During this exchange activity, the core members of our team also chose to listen to the Follow-Leadership lecture given by Brenda, a professor of Berkeley business school. In this lecture, we learned about the qualities we should have as leaders and how to deal with the conflicts in our team. This experience has played a important role in the internal coordination of our team.
Guidance from Predecessors
At the beginning of our project, Li Jiabao, a member of XJTU-China iGEM team in 2018, distributed the teaching video of synthetic biology and sorted out the materials related to biological foundation, which played a great role in the process of project design, and made us have a basic understanding of the commonly used methods of synthetic biology experiment.