Human Practices

Along with developing our project, we conducted an extensive amount of HP activities to further understand the problems we intend to solve and to collect suggestions from professionals and our potential users. In general, our HP work comprises three major sections: 1) A questionnaire-based social study intended to identify the potential cause of stress accumulation in college students, 2) Interviews of synthetic biologists regarding the optimization of our system during the iGEM season, and 3) Interviews of our potential users regarding their ideas and thoughts on how our project might possibly be implemented in the real world.

How our HP work crosstalk with our lab work and science communication program can be seen in the picture below (you can access the detailed information by clicking each box).

1.A questionnaire-based study regardingthe self-awareness of mental stress in college students.

1.1 Rationale

Stress-related mental conditions in academia have raised broad attention for years. In 2018, Evans et al. showed that about 40% of graduate students suffer from anxiety and depression (Evans, 2018). Similar results were also reported by Woolston et al. in 2020 (Woolston, 2020). They showed that more than half of the interviewed postdoc researchers have considered leaving science due to work-related stress, depression, anxiety, and similar issues. In China, Cheng et al. from Fudan University revealed that more than 40% of the reported graduate student suicide cases were related to academic stress, graduation pressure, and depression (Cheng et al., 2020). As a well-educated group of people, college students and young scientists should be knowledgeable enough to be aware of these potentially destructive outcomes caused by chronic stress; we then wondered whether it was the active ignorance of mental stress or the lack of self-awareness regarding the accumulating stress that eventually led to the high incidence of depression and anxiety in this population.

1.2 Methods

To answer our question, we designed a questionnaire to 1) evaluate the self-awareness of the stress levels and 2) understand the stress conditions and how mental stress is commonly dealt with in undergraduate and graduate students. For this purpose, the questionnaire is generally separated into three sections. In the first section, the participants are required to score the mental stress they are currently under on a scale of 0 (totally no stress) to 100 (severely stressed) to provide a quantified self-awareness of stress. In the Second section, the participants are guided through a clinically approved stress assessment scale (Cohen et al., 1983), which provides us with a standardized and quantified approach to evaluating the real stress levels of our participants. In the third section of the questionnaire, we generally ask questions regarding how our participants dealt with mental stress and how our participants evaluated their stress levels. This questionnaire was distributed via the internet to students from National University of Defense Technology, Central South University, and Fudan University with the help of our collaborators.

1.3 Results

We received 690 answers from our participants. Among those, 161 were grad students, and 340 were undergrad students. As expected, we found that only 2.14% of our participants chose to actively ignore the existence of mental stress (Figure 1a), suggesting that most of our participants could actively adjust their lifestyle when they are aware of the increased stress level. Interestingly, when comparing the stress levels obtained by different approaches, we noticed that in both undergrad and postgrad populations, the self-reported stress levels were significantly lower than the score reported by the clinical assessment scales (Figure 1b, P<0.0001, paired Wilcoxon test), suggesting that the population might tend to underestimate their stress levels. Moreover, questions regarding the ways they usually use for pressure measurement revealed that though some students used wearable devices or stress scales to measure their stress, more than 70% of our participants reported that they had never actively measured their stress level.

1.4 Conclusion

Our results showed that although our participants have ways to actively adjust their lifestyles under stress conditions, they generally lacked proper ways to measure their stress levels, which could contribute to the deviation between self-reported stress levels and the levels reported by the clinical assessment scale. To tackle this problem, we decided to build a biological tattoo that changes color when the stress level rises as an alarm for our users.

2.1 Interview with Mirta Viviani

Purpose: Although we’ve demonstrated that a GR_LBD-based Tet_stress can significantly improve GR responsiveness compared to the original GR-based system (see our Results page for more details), we still find the sensitivity of our system unsatisfactory. We’re wondering if it is still possible to improve the performance of the Tet_stress-based circuits. Hence, we approached Mirta Viviani and presented her with our rationale and preliminary data. Mirta is a ph.D. student who works on mammalian cell engineering in the Laboratory of Biosystems Engineering, School of Life Sciences, Westlake University.

What we learned: After we offered a brief introduction to our project, she put forward three potential ways how we can make our system more sensitive:

1. Change VP64, the activating transcription factor we use now, to other transcriptional activating domains like VPR or VP16 on the responsible elements of tetR may help us get higher transcription levels
2. Try different minimal promoters after tetO7 and plasmid ratios.
3. Try different lengths of the linker between the listed functional elements.
4. Manipulating the export or import of the protein via NLS or NES might also help improve our system's efficiency and sensitivity.

Improvements we made:

We adopted her suggestions and performed further linker optimization and NES integration (see our Engineering Success page for details). We finally obtained an optimal glucocorticoid response with the GR_LBD-NES-TetR configuration.

2.2 Interview with Profs. HuijieBian and Ding Wei.

Purpose: After we got the result of the late experiment, we went on to ask how our project could be possibly applied in real-world scenarios. We also wanted to know how to improve our project to meet the clinical needs. Therefore, we interviewed Profs. Huijie Bian and Ding Wei for some discussion. Prof. Huijie Bian is the Deputy Director of the Cell Engineering Research Center/Cell Biology Department of AMMU, which mainly focus on targeted gene therapy. Prof. Ding Wei is the associate professor at the Cell Engineering Research Center who majorly work on oncolytic virus. 

What we learned: After we introduced the project, Professor Bian affirmed our idea, and both she and Professor Wei offered some suggestions about the concrete designs of our project. They also suggested what further research we should take if we want to increase the security and sensitivity of our system.

1. We can conduct further experiments using existing mouse depression models to observe the changes in subcutaneous glucocorticoid content in mice under depressed or high mental tension to adjust the range of our smart cell response to glucocorticoids.
2. We can design the multi-stage cascade reactions; the first level expresses a non-human transcription factor and then import a non-human transcription binding region to avoid initiating the cells’ transcriptional system, which can improve the safety and specificity. At the same time, the multi-stage cascade can also enhance the signal intensity.

Improvement: We decide to prepare the IACUC and apply it for animal experiments to figure out the subcutaneous glucocorticoid concentration under pressure in future work.

3.1 Interview with Zihan Li

Purpose: We consider grad students and young scientists our major end users. To learn more information about their demands and suggestions about our project, we interviewed Zihan Li, a doctoral student at AMMU whose main research areas are stem cell and tissue regeneration.

What we learned: We learned that Ph.D. students might face pressure from lab meetings, lab work, paper submissions, etc., and the pressure tend to accumulate in daily basis. He acknowledged that most of his peers lack proper ways to monitor their stress levels and are willing to use one if there is an inexpensive and convenient solution. After we talked him through our project, Li expressed great interest and also raised some concerns:

1. Safety: The implantation method of smart cells is important, as the size of the wound, and whether it is compatible with other operations (blood draws, surgery, etc.).
2. Privacy: It may not be suitable for showing biological tattoo markers in some public places.
3. Flexibility: We hope the smart cells can be implanted and removed flexibly as needed.

Improvements: 1) We further discussed the pros and cons of different delivery approach (see HP-implementation page for more details); 2) We developed a fluorescent protein-based system to address the privacy concerns (see Proof-of-concept page for details).

3.2 Interview with Dr. Longbiao Cui

Purpose: To determine our project's value and potential application to other potential users and to get more suggestions to improve our project, we interviewed Dr. Longbiao Cui, professor of the Clinical Psychology Teaching and Research Section of AMMU.

What we learned: Mental diseases such as anxiety and depression caused by chronic stress are relatively high (nearly 10%). Our project can also be used as a clinical tool to assist doctors in diagnosing and collecting relevant data for treatment. In the future, it can also be applied to selecting special occupations with astronauts, pilots, and other high psychological state requirements or daily feedback to reduce the non-necessary reduction caused by chronic pressure under special conditions. Dr. Cui also raised some concerns on the aspect of a clinical psychiatrist:

1. The visible stress tattoo itself may become another source of stress.
2. It will be nice to develop a device to record the color changes, allowing doctors to monitor the patient’s stress level in real-time.

Improvements: 1). We developed a fluorescent protein-based system to address privacy concerns (see Proof-of-concept page for details); 2). We plan to characterize the ON-OFF dynamics of our system further to evaluate whether our system can meet the need of clinical psychiatrists. We also plan to design hardware and APPs to more intuitively respond to chronic stress changes through the data