Team:Hangzhou WestLake/Human Practices

Human Practices

Overview

Human practice in the iGEM competition is about introducing synthetic biology to people through various ways. This year, the Hangzhou Westlake iGEM team aims at captivating public interest towards this new field of science by sparking conversations among students in high school and university. We interviewed a number of institutions, including hospitals, environmental protection agencies, and factories, in order to gain in-depth, up-to-date, and contextual knowledge on PCB-induced pollution. Following these practices, we present the information from our iGEM project and the general study of synthetic biology through our Wiki to the public and youth in particular.

What have we done?

Our team has conducted thorough research in areas related to our project. First, we compared aromatic cyclic hydrocarbon substances and determined their toxicity, arriving at the finding that chemical products like paraquat have the same effect as PCBs. To have done so, we interviewed doctors and nurses who had been exposed to PCBs and paraquat poisoning in ICU of the hospital, who warned the public against such exposition to these chemicals. Then we visited insulation oil and lubricating oil factories to learn about the production process and the role of PCB-containing material in production. We also inquired the factory workers specifically on how they avoid the ingestion of PCBs, as well as proper disposal techniques of PCB and discharge production waste (primarily PCB-containing pollutants). To gain deeper understanding, we visited the environmental protection bureau to acquire exact emission data and administrative strategies to alleviate PCB pollution implemented by the government.

To 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 for 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 a detailed treatment and drug description. 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 colorless and odorless high-efficiency herbicide that is soluble in water, paraquat is mortally toxic to living organisms. Research on paraquat holds great significance in the development of human science.

2.Interview with officials at Anhui Province's Bureau of Environmental Protection

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

A: Chinese factories initiated the production of PCBs in 1965 and ceased the process in 1974. In the early 1980s, the total production output was about 1×10^4t, including 9000t of PCB3 and 1000t of PCB5.

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

A: From 1950s to 1980s, China imported a large number of PCB-containing electric capacitors from Belgium, France and other countries without administration permission. Today, the use of most of such equipment has been terminated. Investigations show that PCB content in used capacitor dipping agent is typically above 90%, and PCBs content in used imported transformer oil above 50%. There is limited information on the current import status of raw materials and finished products containing PCBs in China. In incomplete tally, the Tianjin Painting Material Factory in China had produced a type of paint that contained PCBs as early as in the end of the 1950s. China had yet to produce PCBs at that time. Therefore, it was evident that the PCB in the paint were acquired through import.

In addition, I learned from the Chemical Import Company of China that there were records of import of PCBs in 1965. China's import of PCBs-containing materials mainly proliferated in the 1970s and 1980s, when the import of PCBs were primarily intended for the use in special transformers and power capacitors matching with large imported equipment. A small amount of PCBs were imported in the form of hydraulic oil or heat conducting oil. Businesses, such as Liaoyang Chemical Fiber Factory, Wuhan Iron and Steel Co., Shanghai Baoshan Iron and Steel Plant, have all imported PBC transformers and capacitors, mainly for France, Germany, Japan, and Belgium. Further description or detailed report of China's PCBs import would require extensive research and investigation.

Q: What is the contemporary context of PCB use in China?

A: The use of PCBs in China can be divided into three categories: enclosed use (as in lectric power capacitors and transformers), semi-closed use (as in heat conduction oil, hydraulic oil, vacuum pumps, electric switches, voltage regulators, liquid insulated cables, etc.) and open use (as in lubricants, surface coatings, plasticizers, additives, etc.). In practice, PCB3 as power capacitor impregnating agent and PCB5 as paint additive is dominated the use and domestic production of PCB in China. On the other hand, PCBs used in transformers are all acquired through import. Although there is a certain understanding of the use of imported PCBs transformers, the general use of PCB in China is still unclear. Information on PCBs used in other sites is highly 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 having 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 to this project and some of them asked questions after the presentation. Through giving presentations regarding 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 know more about iGEM competition. The aim of the seminar was to let students know more 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 in the future.

3.Our Poster for Public Presentation

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 n current undergraduate student at Cornell University, offered abundant help from the start of the project to the lab execution. As the leader of the HFLS_H2Z_Hangzhou team in 2017, he had a deep understanding of the competition. He attended our weekly meetings and reminded us to keep track of our own work.

Kaixuan Li, a 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. Ziyan Peng was responsible for the final editing and revision 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, Yu Shi, and revised by Ziyan Peng.

Many thanks to everyone who made our iGEM journey amazing!