Team:XMU-China/Human Practices
This year team XMU-China developed cell-free systems to detect and treat diseases. Protein detection is unique and significant in biology fields, especially for the detection of protein biomarkers which produced by diseased cells. In order to overcome the deficiencies of traditional detection methods, we have developed an Aptamer Based Cell-free Detection system (ABCD system) of protein. The core of the ABCD system is the specific binding of the aptamer and its target protein. After protein detection, we use outer-membrane vesicles (OMVs) to treat the diseased cells. We designed a system that has realized the efficient, customizable production of OMVs, which serves to encapsulate specific siRNA for disease treatment. To guarantee the practicability detection and treatment system, we also improved KaiABC system and TDPs system to regulate the expression rate of OMVs and store fragile chemicals or biological materials.
In order to equip our idea with social demands, we have been practicing all through our project by conducting investigation on diseases detection and corresponding information, visiting biotechnology company to gain a better knowledge about the diseases detection, interacting with public to propagate synthetic biology and learning from teams applied themselves in diseases detection theme. We divided these activities in four stages which are Laws and Regulations, Inspiration and Reality, Public and Promotion, Idea and Wisdom, respectively. We have completed Human Practices silver medal criteria, Human Practices gold medal criteria/ Best Integrated Human Practices award and Best Education and Public Engagement award.
In addition，during human practice activities, we found that most disease-diagnosing methods are confined to specific delicate testing apparatus, which are expensive, time-consuming and low sensitivity. The study of Point-of-care testing (POCT), also called bedside testing (with the definition of medical diagnostic testing at or near the time and place of patient care), has become very heated because of its convenience, simplicity and highly efficiency. Internet of things (IoT) is the network of physical devices, vehicles, home appliances, and other items embedded with electronics, software, sensors, actuators, and connectivity which enables the connection and exchange of data.
Here we came up with a design. We combined the idea of our project-Aptamer-based Cell-free Detection system (ABCD system), IoT, and the above concept of POCT so as to develop a microfluidic device, which is small while convenient for real-time detection of cancer. To see the Hardware and XMU-China_Enterprenureship.pdf.
Creating a brainstorm is simplified, how to put theory into practice matters more. For understanding diseases detection reality socially, we visited three organizations, which all have authority background in Xiamen area. In order to consider the feasibility of the ABCD system in our project, we interviewed Prof. ZhiZhu who has very fruitful results on aptamers, the Chief Physician Weiwei Tang and Prof. Zengfu Xue. We finally visited a grass-roots Health Service Center to find our product's market positioning.
On the afternoon of July 9th, accompanied by a line of people，Haifei Xu, the technical director of the Wan-Thai Biological Pharmaceutical Co., Ltd., XMU_CHINA team visited to Xiamen INNOVAX Biotech Co., Ltd., Xiamen InnoDx Biotech Co., Ltd.to study.
The first stop, Xiamen InnoDx Biotech Co., Ltd. According to the company's representative, this company is a wholly-owned subsidiary of the YOSEIDO's Wan-Thai bio-Pharmaceutical Co., Ltd., focusing on the development, production and sales of automatic tubular chemiluminescence immunoassay reagents and bioactive raw materials.
The next stop, Xiamen INNOVAX Biotech Co., Ltd. The same, it is also a wholly-owned subsidiary of the Wan-Thai Bio-Pharmaceutical Co., Ltd., which is committed to providing high quality and affordable innovative vaccines to the public for human disease prevention. Since its inception in 2005, the corporation has successfully listed the world's first vaccine to against HEV.
On September 30th 2018, we visited Mr. Su, founder and chairman of Xiamen Kymem Membrane Technology Ltd, to have a deep discussion about information related to enterprises. He shared with us his experience of setting-up a new company, and rendered us prepared for drawing up business plans.
In the view of Mr. Su, having a sufficient knowledge about not only your company, but your competitors' on the basis of market analysis was the premise of drawing up a good business plan. The business plan should involve some basic parts, namely the company's technology introduction, market potential demand, market actual demand, team composition and competitive team analysis. We introduced our project, which joined competition this year, to Mr. Su, and highlighted the hardware products which we independently developed. Mr. Su expressed his expectation for the potential of our hardware equipment in the medical field. But on the other hand, given that starting a new business is tough, Mr. Su believed that our team would encounter five “new” threats in the initial stage, that is, new team, new cooperation partners, new capital, new technologies and new customers.
He also mentioned that at the initial period of our business, not only the company's internal developing problems, but also the constrictions of the external environment would be the obstacle. Referring to his personal accumulated experience, Mr. Su further gave us some suggestions targeting on starting a new business: considering that China's current intellectual property law system was still imperfect, innovative companies, like ours, who were known for their technological advantages must emphasize the patent-protection. What's more, when our brand was still not so influential compared to those prestigious companies, we should put seeking target customers and broadening sales channels on the front burner, and carefully prevent our technology property right from being infringed. On top of that, he proposed that since our hardware product was based on the "government + grassroots + personal" operation mode, it was necessary to work hard in marketing, with clear targets and strategies.
We visited Mr. Jianghong Wu, president of Xiamen Wenda technology LTD. We had an enjoyed communication with Mr. Wu, which is about the communication mode between team hardware and software and the practicability of software design. We introduced the general situation of this year's team project to Mr. Wu, and emphasized the original intention of the team to solve the "aerial phenomenon" of the current "graded diagnosis and treatment" policy, as well as the difficulty in getting medical treatment in remote areas. Mr. Wu fully affirmed the innovative project and praised the team consciousness of social responsibility.
As for APP part he gave some professional advices and technical guidance, like how to accelerate our convolution neural network recognition speed with collected increasing number of images cases, and how to link block them up between chain technology public health service database to serve the public better. In addition, Mr. Wu gave us a lot of advice on product promotion and profit model.
In order to consider the feasibility of the ABCD system in our project, we interviewed Prof. Zhi Zhu of the College of Chemistry and Chemical Engineering of Xiamen University on May 2, and she has very fruitful results on aptamers.
The professor affirmed the feasibility of our project and gave us some invaluable advice. Under the suggestion of Prof. Zhi Zhu, we chose EpCAM and aptamer SYL3C. In addition, we are very grateful to Prof. Zhu for her support on many instruments and materials provided during our experiments.
IThis year，team XMU-China developed new methods named ABCD System and OMVs Treatment for detecting and treating diseases. In order to prove the feasibility of our project, we consulted the Chief Physician Weiwei Tang in Medical Oncology, The First Affiliated Hospital of Xiamen University, on August 23th. We introduced briefly the structure and function of our self-designed hardware Fang to Dr. Tang. During the talk, Dr. Tang appreciated it that our ideas are innovative and helpful. What's more, Dr. Tang also told us those frequently-used and classical clinical detection methods, like ELISA, CLIA and so on. Most of which are time-consuming, expensive and complex, compared with Fang. As for Fang, Dr. Tang agreed with our proposals that we are going to make Fang widely used among basic medical institutions, especially in remote, and poverty-stricken areas because of its convenient and practicability. But it's worth noting that today there is an authoritative method for disease-diagnose, i.e., pathological diagnosis. So our detection method would just serve as a kind of screening tool rather than diagnosis method, according to Tang's suggestion.
The professor affirmed the feasibility of our project and gave us some invaluable advice. Under the suggestion of Prof. Zhi Zhu, we chose EpCAM and aptamer SYL3C. In addition, we are very grateful to Prof. Zhu for her support on many instruments and materials provided during our experiments.
We visited Prof. Zengfu Xue, deputy director of the Cancer Early Screening Diagnosis and Treatment Center, the First Affiliated Hospital of Xiamen University. We exchanged and discussed with Prof. Xue about the design of cell free detection system of tumor markers and detection carrier of microfluidic chip. Prof. Xue fully affirmed the innovation and feasibility of this project, and appreciated the original intention of the team to solve the problem of difficult access to medical services in large hospitals, which helped to improve the current "hierarchical diagnosis and treatment" policy system to some extent. We communicated with Prof. Xue about the next clinical trial plan of the detection hardware Fang, and reached a preliminary cooperation agreement. If possible, we could take the instrument to the lab in hospital, and use the patient's blood sample to verify the performance of the instrument, such as stability and sensitivity.
We visited Health Service Center of Datong Street, Tong'an District, Xiamen city. We learned about the development of tumor markers detection work in primary hospitals from director Zhan. We found that primary hospitals were mostly able to carry out screening of basic tumor markers such as alpha fetoprotein. If people want to carry out more systematic and specific tumor screening, they had to be referred to higher-level hospitals. In addition, due to the implementation effect of China's medical security system and Xiamen's health policy, the screening of tumor markers at the grassroots-level health center could save about 50% of the cost compared with the large hospitals. Therefore, to sum up, president Zhan affirmed the future prospects of our instruments, believing it would solve problems such as "difficult access to medical services" in real life, to better provide more convenient medical and health services.
The medical reform policies, together with the reasonable superintendence will promote the rapid progress of POCT（point-of-care testing） in America. [1]
America now is encountered with the payment pressures imposed by enormous medical insurance fees resulted from the aged tendency of population and the changes in the spectrum of diseases. According to the forecast by CBO, the expense of health products will reach up to 4,600 billion in 2020, accounting for 20% of the GDP. Besides, it was shown in the OECD report that in 2011, American people's average expenditure on health care was up to 8233 dollars, twice the expenditure of comparatively developed European countries like French, Switzerland and England. In March, 2010, American president Barack Obama carried out the “Affordable Care Act”, aiming at improving the quality of American health insurance service, and decreasing the cost of health care service. The “ACA Act” established the balance sharing plan, which encouraged doctors and hospitals to meet the patients' medical needs by an integral medical service system through encouraging accountable care organizations. Under that balance sharing plan, suppliers and organizers of the service who meet the standard set by US HHS can serve those who benefits from the medical insurance plan through co-organizing and coordinating. ACO could help reduce the cost and increase the efficiency, and distribute part of the surplus among members. Under the design of the ACO system, the new service supplying mode integrate hospitals, general practitioners, medical specialists and other specialists to answer for the medical quality and medical cost together. [2]
In America, POCT is supervised by FDA, and is co-organized by certification authority, POCT committee, and POCT coordinators. It exists as one of the branches of IVD. Therefore, it is also supervised under the policies of medical apparatus and instruments. At the same time, IVD products are also supervised under CLIA 88 enacted in 1992, which divided IVD into three registering levels (low, medium and high) basing on the complexity of experiments. Before obtaining the right of market entrance, producers have to make appliances according to CLIA categorizing system. After that, CMS, DCLD and CDC will check the products and authorize corresponding CLIA certifications. When it comes to POCT products specially, they will have to firstly recognized by JCAHO and CAP, and finally authorize and certificate by JCAHO, CMS and CAP. [3]
In terms of the usage of POCT products, owing to the fact that there is not much barrier, the superintendence is therefore the key to optimizing POCT and enlarging its range of application. POCT committees have been set in every organization, city and state in America so as to implement quality-superintendence over its usage and guarantee the accuracy of the testing results. [4]
References
[1] https://www.house.gov/
[2] https://www.aha.org/
[3] http://www.gf.com.cn/
[4] https://www.fda.gov/
Working on synthetic biology in a lab is amazing, but sharing our enthusiasm can be even more wonderful. For the science education, we worked with people from all walks of life including teenagers, citizens, freshmen and XMU students.
This year our team XMU-China came to Xiamen Science and Technology Museum to advocate the synthetic biology and our project. The whole project about 3 parts, game, Poster and Wish Wall. First, the game part, children play the roles of antibody and antigen respectively, they battled to be the winner by RPS. After the game they all left a deep impression of the general method that our body fight with cancer. Second，Poster，we had some talkative teammates to take charge of this part, so many student from high school and primary school were attracted ,they crowded in front of the poster and listened to our announcers absorbedly. Finally, the Wish Wall part, after our diverse introductions about the synthetic biology, they had learn a lot about it, and some of them even inspire to give their contributions to synthetic biology, and want to attend the innovative, fantastic competition —— iGEM. It's also a cheerful Day for our team to propagate the synthetic biology to so many children.
Fighting for Mooncake is a traditional activity spent with relatives or close friends in Xiamen area in order to celebrate the Mid-Autumn Festival. Throughout the game, we advertised the iGEM to freshmen, and introduced our project this year. It was worth mentioning that we also collected solicited opinions from all the students about the feasibility of our disease detective device and treatment device with cell-free system this year. 86.7% of participants showed positive attitudes towards our detective device, with the common sense of this device could performed pre-diagnosis quickly to guide patients. 66.8% students thought that it was difficult to treat tumors with vesicles, but it was very promising.
In order to popularize the common knowledge of biology to the public and meanwhile to draw the public attention to synthetic biology, this year we XMU-China team visited to Yanwu Elementary School, volunteering to teach elementary students biology, introducing our team and work, and directing them to do some simple biological experiments. For instance, we let them extract DNA from bananas.
In the course of our introduction, the children of Yanwu Primary school expressed their great enthusiasm, responded positively to our interaction, and moreover they showed terrific zest for our introduction. In these next simple experiments, the children were thrilled to be involved in one of the experiments.
In the end, through our teaching and guidance, most of the children have achieved remarkable results during the experiments. After the event, Yanwu Elementary school students said they learned a lot of useful knowledge, and very happy to do these interesting experiments by themselves within our guidance.
Also, the teacher fed back that they think this activity is so meaningful, extremely hoping to hold more such events in the primary school campus.
No one can make great progress without advice and golden ideas from their excellent partners. During last six months, we participated in some iGEM conferences or even worked as the innovative online meeting organizers. Hopefully these events contributed greatly to all of us.
The 5th CCiC, which full name is the Conference of China iGEMer Community Meetup, was held at ShanhaiTech University on the subject of “Among us, past, present and future meeting” from August 27th to 31th. Dozens of teams from all over the China as well as the Executive Vice President and Chief Operating Officer if the iGEM Foundation, Meagan Lizarazo took part in. During this conference, we fortunately got the chance to have deep communication with many iGEM teams including iGEM_BIT(Beijing Institute of Technology University), iGEM Tianjin (Tianjin Unniversity) and LZU_China (Lanzhou University), and here we really appreciated their advices on our project about the competitive experiments, Kai ABC systems and the OMVs systems. Luckily, we won the best presentation award. What's more, we were greatly benefited from Meagan's lecture pointing out the importance of “After iGEM”. This strengthened our determination to contact the old members of XMU-China and luckily we got precious experiences and further understanding about this competition.
From July 30th to August 3rd, 2018, we participated in the 6th Asia-Pacific iGEM Conference hosted by National Chung Hsing University, Taichung. There was a total of 22 teams and 140 participants from Taiwan, China, and Japan, warming up for the final Jamboree in Boston at the end of October.
The conference invited wonderful speakers from the field of Synthetic Biology and Ambassador of the iGEM HQ to give a talk at the Opening Ceremony. Among them, Prof. Kenji Tsuge from Kobe University explained the emerging synthetic biology technology that can link quantities of large fragments of genes and perform these genes; Dr. Zhang Jui-Jen, an associate researcher at China Medical University Hospital, shared his experience in using synthetic biology entrepreneurship; iGEM Asian Ambassador Chen Hong explained some new systems and shared experience on behalf of iGEM. We gained tons of new ideas from these lectures, which had great influences on our next work.
In the five-day conference, we also had a 20-minute English presentation and a poster exhibition. It was the first time to present our project frankly to iGEMers from Asia, and a good opportunity to practice for the final Jamboree. At the end of the exhibition, Prof. Wenming Yang from the field of molecular biology at National Chung Hsing University gave a comment on our presentation. Through the conference, we also made suggestions to other Asian teams, longing for more cooperation opportunities.
This conference has provided us tremendous new ideas to improve our project and final presentation.
Collaborations with other teams are indispensable links in the iGEM competition. You can click here to see what collaborations that we have done this year, and what achievements we have got.
This year, Team XMU-China is devoted to combine WET LAB, DRY LAB and Human Practices to form an organic integration, while the Human Practices function as a significant theme. As Peter Carr said, "Human Practices is the study of how your work affects the world, and how the world affects your work." Based on this saying, our team will show the inspiration of our project, the ideas of how to design Human Practice activities and how the activities influence our project in return. What we can feedback to the society is also the point we'll show below.
Before having the discussion with Prof. Zhi Zhu, we read related literatures [1], and made the initial determination of using the aptamer sequence, SYL3C, which was referred to in literatures. In the beginning, we came up with three methods (Figure 1) to realize the ABCD system. However, we had no idea about the concrete operations. That was just the reason why we had the meeting with Prof. Zhi Zhu. After listening to the three methods proposed by us, Prof. Zhi Zhu offered personal opinions about them. After taking time restriction and operation feasibility into considerations, we ultimately chose the “competing method” (Figure 1.B) to realize our system. Not only did Prof. Zhi Zhu give us beneficial recommendations, but gave us specific guidance about how to apply the “competing method”. For example, he proposed that we had better use streptavidin magnetic beads as the carriers, and attached biotin on the end position of SYL3C aptamer sequence.
Figure 1. Initiative methods to enable ABCD system. A) Method A. B) Method B. C) Method C.
Therefore, the close combination of streptavidin and biotin could be exploited to fix the aptamer. She also reminded that in order to keep the reactions of competition as far away as possible from the surface of the carriers, TEG could be inserted between biotin and the aptamer sequence. In that case, TEG could serve as a spacer to reduce the effects of reactions on the surface. Suggestions like these, which were offered by our professor were quite a lot. These suggestions enormously facilitated the design of our experiment. On top of that, professor Zhu also listened to our ideas about the the designed model of thermodynamics (See more details in our Modeling page). From her perspective, our ideas were quite acceptable. What more, she pointed out that we could try the “Complementary Trand”, which uses 9nt or even more basic groups, to realize the mutual complement. This suggestion also benefited our experiment and modeling to some extent. Another case worth mentioning is that, when we were encountered with difficulties when using flow cytometry to verify the combination of SYL3C and EpCAM, it was professor Zhu who helped us out of the woods. She suggested us to use another method, fluorescence polarization, to verify the combination of those two molecules [2]. Her advice turned out to be wise and feasible. Finally, we used that method and successfully verified the combination. (See more details in our Results page).
After Fang's sketch completed
In order to seek confirmation about the prospect of our project and Fang on cancer detection, we consulted this on Prof. Xue. We exchanged and discussed with Prof. Xue about the design of aptamer-based cell free detection system (ABCD System) and the design of microfluidic chip drives by centrifugal force. On the one hand, Prof. Xue fully affirmed the innovation and feasibility of this project. On the other hand, Prof. Xue though it that if we could find a more specific biomarker from one cancer like liver cancer, which would strengthen the academic persuasion of our project. We communicated with Prof. Xue about the next clinical trial plan of the detection hardware, and reached a preliminary cooperation agreement. If possible, we could take the instrument to the lab in hospital, and use the patient's blood sample to verify the performance of the instrument, such as stability and sensitivity.
After the first generation product of Fang completed
In order to prove the feasibility of our project, we consulted the Chief Physician Weiwei Tang. During the talk, Dr. Tang appreciated it that our ideas are of social significance. What's more, Dr. Tang also told us most of those frequently-used and classical clinical detection methods, like ELISA, CLIA and so on are time-consuming, expensive and complex, compared with Fang.
As for Fang, Dr. Tang agreed with our proposals that we are going to make Fang widely used among basic medical institutions, especially in remote and poverty-stricken areas because of its convenient and practicability. But it's worth noting that today there is an authoritative method for disease-diagnose, that is pathological diagnosis. So our detection method would just serve as a kind of screening tool rather than diagnosis method, according to Tang's suggestion. We also should pay attention to the testing sensitivity of our project if it could meet national standards or even better.
After the first generation of software tool completed
In order to improve our software part, we had an enjoyed talk with Mr. Wu about the communication mode between team's hardware and software, especially practicability of software design. Mr. Wu fully affirmed its innovation and praised team's consciousness of social responsibility. Also he gave us his own opinions on our project. As for APP part, he gave some professional advices and technical guidance, like how to accelerate our convolution neural network recognition speed with collected increasing number of images cases, and how to link block them up between chain technology public health service database to serve the public better. It's key point for us to develop and improve our software that we want to combine specific functions with advanced tool like AI. In addition, as an outstanding entrepreneur, Mr. Wu gave us a lot of advice, from his early experience, on product promotion and profit model.
After the final generation product of Fang
As we imaged, we want to apply our hardware among grass-root medical center in the future. So we visited Health Service Center of Datong Street to learn about the development of tumor markers detection work in primary hospitals from President Zhan. We found that primary hospitals were mostly able to carry out screening on only one kind of basic tumor markers such as alpha fetoprotein (AFP). If people want to carry out more systematic and specific tumor screening, they had to be referred to higher-level hospitals. What's more, due to the implementation effect of Chinese medical security system and Xiamen's health policy, the screening of tumor markers at the grassroots-level health center could save about 50% of the cost compared with the large hospitals, which is beneficial for people to have cancer detection on regular. It also met our vision about the social role of our hardware. Therefore, to sum up, president Zhan affirmed the future prospects of our instruments, believing it would solve problems such as "difficult access to medical services" in real life, to better provide more convenient medical and health services.
Before we design the Business Plan
In order to learn about startup and operate a company in medical field, we visited Mr. Su. He shared with us his experience of setting-up a new company, and rendered us prepared for drawing up business plans.
In the view of Mr. Su, having a sufficient knowledge about not only your company, but your competitors' on the basis of market analysis was the premise of drawing up a good business plan. Given that starting a new business is tough, Mr. Su believed that our team would encounter five “new” threats in the initial stage, that is, new team, new cooperation partners, new capital, new technologies and new customers. Referring to his personal accumulated experience, Mr. Su further gave us some suggestions targeting on starting a new business: considering that China's current intellectual property law system was still imperfect, innovative companies, like ours, who were known for their technological advantages must emphasize the patent-protection. What's more, when our brand was still not so influential compared to those prestigious companies, we should put seeking target customers and broadening sales channels on the front burner, and carefully prevent our technology property right from being infringed. On top of that, he proposed that since our hardware product was based on the "government + grassroots + personal" operation mode, it was necessary to work hard in marketing, with clear targets and strategies.
Working on synthetic biology in a lab is amazing, but sharing our enthusiasms can be even more wonderful. For the science education, we worked with people from all walks of life including teenagers, citizens, freshmen and XMU students.
You can see more details by click here.
Laws regulate the human world steadily and sustainably, without which we may fall into disorder. The medical reform policies, together with the reasonable superintendence will promote the rapid progress of POCT（point-of-care testing） in America [3].
America now is encountered with the payment pressures imposed by enormous medical insurance fees resulted from the aged tendency of population and the changes in the spectrum of diseases. According to the forecast by CBO, the expense of health products will reach up to 4,600 billion in 2020, accounting for 20% of the GDP. Besides, it was shown in the OECD report that in 2011, American people's average expenditure on health care was up to 8233 dollars, twice the expenditure of comparatively developed European countries like French, Switzerland and England. In March, 2010, American president Barack Obama carried out the “Affordable Care Act”, aiming at improving the quality of American health insurance service, and decreasing the cost of health care service. The “ACA Act” established the balance sharing plan, which encouraged doctors and hospitals to meet the patients' medical needs by an integral medical service system through encouraging accountable care organizations. Under that balance sharing plan, suppliers and organizers of the service who meet the standard set by US HHS can serve those who benefits from the medical insurance plan through co-organizing and coordinating. ACO could help reduce the cost and increase the efficiency, and distribute part of the surplus among members. Under the design of the ACO system, the new service supplying mode integrate hospitals, general practitioners, medical specialists and other specialists to answer for the medical quality and medical cost together. [4]
In America, POCT is supervised by FDA, and is co-organized by certification authority, POCT committee, and POCT coordinators. It exists as one of the branches of IVD. Therefore, it is also supervised under the policies of medical apparatus and instruments. At the same time, IVD products are also supervised under CLIA 88 enacted in 1992, which divided IVD into three registering levels (low, medium and high) basing on the complexity of experiments. Before obtaining the right of market entrance, producers have to make appliances according to CLIA categorizing system. After that, CMS, DCLD and CDC will check the products and authorize corresponding CLIA certifications. When it comes to POCT products specially, they will have to firstly recognized by JCAHO and CAP, and finally authorize and certificate by JCAHO, CMS and CAP [5].
In terms of the usage of POCT products, owing to the fact that there is not much barrier, the superintendence is therefore the key to optimizing POCT and enlarging its range of application. POCT committees have been set in every organization, city and state in America so as to implement quality-superintendence over its usage and guarantee the accuracy of the testing results [6].
References
[1] Yanling Song, Zhi Zhu, Yuan An, Weiting Zhang, Huimin Zhang, Dan Liu, Chundong Yu, Wei Duan, Chaoyong James Yang, Selection of DNA Aptamers against Epithelial Cell Adhesion Molecule for Cancer Cell Imaging and Circulating Tumor Cell Capture, Anal Chem, 2013, 85, 4141-4149.
[2] Liang Cui, Yuan Zou, Ninghang Lin, Zhi Zhu, Gareth Jenkins, Chaoyong James Yang, Mass Amplifying Probe for Sensitive Fluorescence Anisotropy Detection of Small Molecules in Complex Biological Samples, Anal Chem, 2012, 84, 5535−5541.
[3] https://www.house.gov/
[4] https://www.aha.org/
[5] http://www.gf.com.cn/
[6] https://www.fda.gov/
