Team:Hangzhou WestLake/Entrepreneurship

Human Practices

Overview

Human practice in the iGEM competition is about introducing synthetic biology to people through various ways. This year, Hangzhou_Westlake iGEM team wanted to arouse people’s interest towards this new field of science by organizing some talks for both middle school and high school students. We also interviewed several institutions such as hospitals, Environmental protection agencies and factories to learn in-depth knowledge and current situation about PCBs pollution. Through these activities, we presented the basic information about iGEM and synthetic biology to the youth and the public.

What have we done?

We did promotions and researches from all aspects that relate to our project. We first compared aromatic cyclic hydrocarbon substances and determined their toxicity, and found that chemical products like paraquat have the same effect as PCBs. Therefore, we interviewed doctors and nurses who had been exposed to PCBs and paraquat poisoning in ICU of the hospital, and also received warnings from doctors and nurses to the public. Then we went to the insulation oil and lubricating oil factory to learn about the production process of insulation oil and the role of materials containing PCB in the production. We also asked the factory workers how to avoid PCB ingestion and how to dispose of and discharge production waste (including PCB pollutants). We even went to the environmental protection bureau for investigation to get the exact emission data, and how the government planned to deal with the most polluted areas.

For public engagement, we held seminars on synthetic biology and its applications, and hosted exhibitions of our projects on PCBs detectors in the community and in each team member’s high school. We talked to audiences who eagerly concerned about the safety and potential of synthetic organisms including common residents, teachers, students, investors and engineers. We have established long-term relationships with a number of African land resource agencies. We also work closely with environmental protection agencies and city management agencies to show how we can combine microbial plants to control pollution emissions and address PCB emissions. Through direct interviews, in-depth conversations, and presentations, we learned about their concerns and how to embrace the design that is relevant to synthetic biology better. In addition, we also took some videos to show the public how to use our detector and upload them to popular media platforms.

What have we learned?

We believe science is everywhere in society and it is a part of each person’s everyday life. For most people with no science background, synthetic biology seems to be a very difficult topic for them. The take home message: for many, synthetic biology is unfamiliar and unless careful with wording — frightening. It is exceptionally important and too often given too little weight in science: communication and public engagement. Creating, organizing and participating in public events is what has made the real biological application — before it ever was.

Integrated Human Practices

1.Online Conference with an ICU doctor who studied aromatic cyclic hydrocarbon chemicals related diseases

On August 28th, 2019, our team had a skype conference call with Dr. Li, a current ICU doctor in *** Hospital. In her career, she met a lot of patients poisoned by aromatic cyclic hydrocarbon chemicals such as PCBs and paraquat. We interviewed her for her study and experiences with dealing these diseases and suggestions and warnings to common people. The following is the complete record of the conference:

Q: How to deal with paraquat poisoning in the first time? What is the method of treating paraquat poisoning?

A: Take mud water nearby. If there is no clean mud water, take two packs of Smecta to absorb toxins. Immediately go to the nearest hospital for gastric lavage.

Q: How does paraquat cause death?

A: Acute respiratory failure or pulmonary fibrosis for 7-14 days. The lethal mechanisms include multiple organ failure, oxidative stress and DNA damage caused by the outbreak of inflammatory factors.

Q: Are there any relevant cases to share?

A: There are many cases of suicide by drinking paraquat, and there are cases that paraquat is drunk by accident. The situation is always that farmers mistake the paraquat that is forgotten in the beverage bottle as water while working; or working from the farmland, returning home, and thirsty, the farmers drink the paraquat by accident carelessly.

There are also many cases of eating poisoned vegetables that have just been sprayed with paraquat. We have treated many cases of paralyzed amaranth and other green vegetables, which are often treated by a family. There is also a part of the farmers who mix and dilute the paraquat in agitation to inhale the skin and contact with paraquat. Our department admitted a case of agitating pesticides by hand this month, and contacted the scrotum by hand. After 10 days, the farmer came to our hospital for rupture of the scrotum, and died of lung disease and respiratory failure.

Q: How do you usually take care of such patients?

A: Nursing is also very important. Oral ulcers, local skin ulcers need to be particularly nursed. Psychological care for most patients with paraquat poisoning is more important.

Q: How many cases will there be in a year?

A: The number of cases in the largest hospital in Henan Province is about 300 a year.

Q: Do you have anything to tell everyone about paraquat?

A: Yes, let me provide to a detailed treatment and drug desription. Regarding the treatment of paraquat, and the 2013 expert paraquat consensus. The specific treatment of our hospital is as follows: The treatment plan for acute paraquat in our hospital is in accordance with the accepted treatment plan of the expert consensus on acute paraquat poisoning recommended by the Emergency Physician Branch of the Chinese Medical Association, but our specific details are more practical and optimized. Specifically:

• (1)gastric lavage. All patients were enrolled in a single lavage of room temperature water, and the gastric lavage was terminated after clearing the gastric lavage. Whether or not the patient has been lavaged in the local area before coming to our hospital, the patient has to lavage in our hospital after entering our hospital.
• (2)Drug application.

1. After gastric lavage, give Smecta (montmorillonite powder) 6 g (2 packs) plus 50 ml of water and mix orally, once every 4 to 6 hours.

2. After Smecta oral administration, 20% mannitol was administered orally in 100 ml orally, 4 to 6 hours, and alternately applied with Smecta (montmorillonite powder). Maintain for 3 to 5 days. Because PQ patients have severe oral ulcers in almost all patients after 2 to 3 days of taking poison, it is difficult to drink water, and it is difficult to take oral drugs at this time, and the use of Smecta and mannitol is stopped. During the period using Smecta, the color of the stool is closely observed. Generally, the green color indicates that the intestine still retains paraquat. To continue taking Smecta orally, give 20% mannitol 100 ml orally, 4 to 6 hours once, with Smecta (Alternate application of montmorillonite powder), if the stool turns yellow, showing normal traits, you can stop taking Smecta.

3. Furosemide needle, intravenous bolus injection, 40 mg / time, 2 times / day. Pay attention to the electrolyte balance when using, supply calcium and potassium to patients.

4. Methylprednisolone 15mg/ (kg · d), reduce the amount after one week, reduce the amount every three days, reduce 40mg each time; (informed consent for drug side effects with patients or their families)

5. Cyclophosphamide, 10-15 mg / (kg · d), stop using when used for a week; (informed consent for drug side effects with patients or their families) can be discontinued if bone marrow transplantation or renal dysfunction occurs.

6. In order to avoid calcium deficiency and osteoporosis caused by glucocorticoids, calcium supplementation and potassium supplementation are given to patients. Calcium supplementation commonly used calcium carbonate D3 oral 0.75g (equivalent to calcium 300 mg, vitamin D360 international unit), and is recommended to use nasal Calcitonin Nasal Spray nasal spray, the amount is 200IU / (spray · d), Potassium supplementation tablets 0.5 po TiD were applied to potassium supplementation. We will check the genetic determination of femoral head necrosis caused by hormones. If it is highly dangerous, we will use general sodium phosphate to prevent femoral head necrosis.

7. In order to avoid stress ulcer caused by glucocorticoids, the patient proton pump inhibitor omeprazole, 40mg, bid intravenous drip.

8. To avoid infection with a low immune status caused by the use of large amounts of hormones and immunosuppressive agents, azithromycin is used to combat infection. Azithromycin has an anti-pulmonary fibrosis effect. Usually 0.5g intravenous drip × 5 days, antibiotics with small side effects such as penicillin can be used prophylactically. Once there is definite evidence of infection, then target the antibody to escalate, and take specimens from the appropriate sites for pathogenic examination.

9. Propranolol, which reduces the amount of PQ that accumulates through the dopamine system to the alveolar epithelium. Using Propranolol 10-20mg 3 / day can compete with PQ combined in lung tissue. Maintain heart rate no less than 50 times / min, continue to take until the patient is cured.

10. Bailing capsules, 0.5tid.po, taken daily. After the patient discharged from the hospital for one month, review chest CT, if there is no obvious abnormalities in the lungs, the medicine can be discontinued.

11. Xuebijing injection, 50mg-100mg/day, should be discontinued if there is bleeding tendency. Three days before admission, heparin is often used in blood purification to prevent thrombosis due to hemoperfusion. At this time, hemagglutination is poor and there is a tendency of bleeding. At this time, no "Xuebijing" is needed.

12. Edaravone, 30 mg / day, can scavenge oxygen free radicals, but those with kidney damage are deactivated or not used.

13. Ulinastatin, 30 mg / time, 2 times / day, intravenous drip, can treat multiple organ failure.

14. Reduced glutathione, 2.4g / day, intravenous drip, detoxification, liver protection.

15. Compound glycyrrhizin, 120 mg / day, liver protection, regulation of immunity.

16. When the patient has cough, apply doxofylline 0.3 / day, oral or intravenous drip, ambroxol 90mg / day, oral or intravenous drip.

17. Patients with oral ulcer and tongue erosion were treated with Kangfuxin liquid orally and swallowed three to four times a day, and Fuji Re-spray (for external use of human recombinant fibroblast factor) three to four times a day.

• (3) Blood purification.Hemoperfusion (HP) is performed as soon as possible after admission. In general, the whole process of hemoperfusion can be completed quickly within one hour, such as talking with patients' families, signing informed consent, central venous catheterization and on-line hemoperfusion. Hemoperfusion lasts for 4 hours, once a day, at least three times in a row, and then stops. If the patient has acute renal failure, hemodialysis is performed. Informed consent is signed by each patient or his family member for hemoperfusion.
• (4) Daily measurement of PQ concentration in urine. The specific method of urine PQ determination: urine PQ concentration by 722 spectrophotometer, using sodium bicarbonate, sodium carbonate, sodium dithionite as a color developer, applying chemical colorimetry. The normal reference value is (0-0.2) ug/ml.
• (5) Oxygen therapy. The patient is routinely not taking oxygen. Only when the oxygen saturation is less than 70% or the arterial blood gas analysis oxygen partial pressure is less than 50%, and is accompanied by symptoms and signs related to respiratory distress.

Q: What stage are scientific researches towards aromatic hydrocarbon lead diseases on?

A: Scientific research on paraquat has been ongoing. Every year, the National Natural Science Foundation publishes many SCI papers and Chinese core journals. As a water-soluble colorless and odorless high-efficiency herbicide, paraquat has a high mortality rate, and its research has great significance for the development of human science.

2.Interview with staff working in Anhui Environmental Protection Bureau

Q: What is the production of PCBs (raw materials) in China?

A: China began to produce PCBs in 1965, and most factories stopped producing them in 1974. In the early 1980s, the total production was about 1×10^4t, including PCB39000t and PCB5 1000t.

Q: What is the import history of PCBs in China?

A: From 1950s to 1980s, China imported a large number of electric capacitors containing PCBs from Belgium, France and other countries without being informed. At present, most of these equipment has been scrapped. According to the investigation, PCBs content in used capacitor dipping agent is more than 90%, and PCBs content in used imported transformer oil is more than 50%. There is little information on the import status of raw materials and finished products of PCBs in China. According to incomplete statistics, Tianjin Painting Material factory in China had produced a kind of painting material that contains polychlorinated biphenyls (PCBs) at the end of 1950s. China did not produce polychlorinated biphenyls at that time, so it is the first polychlorinated biphenyls that China imported as raw material.

In addition, I learned from China's chemical import company that there was a record of import of PCBs in 1965. China's imports of PCBs-containing equipment were mainly concentrated in the 1970s and 1980s, when the imports of PCBS were mainly concentrated on special transformers and power capacitors matching with large imported equipment, and a small amount of PCBS were imported in the form of hydraulic oil or heat conducting oil. Such as Liaoyang chemical fiber factory, Wuhan iron and steel company, Shanghai Baoshan iron and steel plant, have imported polychlorinated biphenyl transformers and capacitors, imported countries mainly for France, Germany, Japan, Belgium. The detailed description or report of China's PCBs import needs further work.

Q: What are the usage conditions of PCBs in China?

A: The use of PCBs in China can be divided into three ways, namely the enclosed use (used in the electric power capacitor and transformer), semi-closed use (such as heat conduction oil, hydraulic oil, vacuum pump, electric switch, voltage regulator, liquid insulated cable, etc.) and the open use (such as lubricants, surface coatings, plasticizers, additives, etc.). According to the actual use of China's PCBs, the use of PCB3 as the power capacitor impregnating agent and PCB5 as the paint additive is relatively clear, which is the main use field of China's PCBs, and the domestic PCBs are basically used in this aspect. PCBs used in transformers are all imported. Although there is a certain understanding of the use of imported PCBs transformers, the overall situation is still unclear. Information on PCBs used in other sites is very limited and, if used, the source may be mainly imported, but the use will not be large and will require further verification and investigation.

Q: What is the storage condition of PCBs in China?

A: From 1980s to early 1990s, PCBs power capacitors were stored in China. According to the production and use life of PCBs capacitors, this is the peak period when PCBs capacitors are out of use in China, and it is also the period when PCBs storage is the largest in China. However, as the storage was carried out by the region or the enterprise alone, there was no sound and effective reporting system. According to the “National Inventory Report on PCBs Containing Polychlorinated Biphenyls” completed by CEC in 2006, the situation varies greatly from place to place. At the same time, in some areas, according to the "Provisions for the Prevention of Environmental Pollution of Polychlorinated Biphenyl-Containing Electrical Installations and Their Wastes", the electrical equipment containing PCBs was sealed. The qualified areas and units carried out centralized storage of caves, and some areas used iron boxes. After storage, it will be stored in the idle warehouse or the open space of the factory.

Public Engagement

1.Presentation delivered to Middle School Students

On Sep 10th, we made a presentation in front of 50 middle school students who were now have Advanced Science Lab courses. The presentation consisted of an introduction about iGEM, synthetic biology and our project --- PCBs detector. Since some of the audiences were still in 7th grade, we used many cartoons and analogical examples to illustrate the basic concept such as BioBricks and synthetic biology. Most students showed their interests in this project and some of them asked questions after the presentation. We believe that giving presentation about iGEM to younger students, we can inspire them to think deeply about how synthetic biology can change our lives.

2.Seminar for Potential Biochemists/ Bio-engineers in High School

On Sep 24th, we had another seminar for high school juniors who are interested in BME major or Biochemistry major. We also prepared some flyers about our project so they can have deeper understanding about iGEM competition. The aim of the seminar was to let students have better understanding about the applications and benefits of synthetic biology. At the same time, we wanted to arouse their interests in this relatively new scientific field and encourage them to join iGEM competition in the future.

3.Our Poster to Public

Attributions

Hangzhou_Westlake iGEM 2019 would like to express thanks to the following people and organizations for their contribution to our team:

Academic Guidance

Dr. Pui Yan Ma, the primary instructor of the team and the research fellow at Harvard Medical School, helped us with the brief design of the project and our experiments.

Dr. Jie Yan, the secondary instructor of the team and the biology professor at Zhejiang university, taught us the basic process of conducting scientific research and collecting data. He also helped us out with troublesome experiments.

Jianan Li , the advisor of the team and an current undergraduate student in Cornell University, offered abundant help from the start of the project to the lab execution. He was a former HFLS_H2Z_Hangzhou and therefore he was proficient in He attended our weekly meetings and reminded us to keep track of our own work.

Kaixuan Li, an current graduate student in Zhejiang Univerisity, help our team member with Wet Lab experiments.

Cooperation

ZJU iGEM 2019 mentored our team during the summer. ZHJ Medical School provided us an advanced wet lab. Our sponsor, Hangzhou Punuo Education ., Ltd helped our team with public engagement.

Project Design & Wet Lab Experiment

Jiayue Liu and Jianan Li did the basic project design under the help of Dr. Pui Yan Ma. Xinchen and Zhenhe did most of the wet experiments, including biobrick design, part assembly and validation scheme design.

Human Practice

The intergrated human practice is done by Jiayue Liu and Yu Shi. The presentation were organized by Jiayue Liu and Yu Shi. Jiayue wrote an article about our project and interviewed the environmental protection agencies and factories. Wenyu Shi interviewed the ICU doctors in the hospital.

Graphic Design

Jiayue Liu did the poster and Logo design of our team. Pengziyan was responsible of the graphic design of our Wiki page.

Wiki Construction

Wiki was constructed by Letian Tao, who learned web design starting from scratch in this summer. All the contents on the Wiki were written by Jiayue Liu, Xinchen Lyu and Yu Shi.

Many thanks to everyone who made our iGEM journey amazing!