<!DOCTYPE html> Safety

SAFETY

Introduction

Safety, which is an eternal topic of social concern, is the prerequisite for mankind to understand and transform the world, so our project is no different.

First of all, what is safety? Safety is to control the possible damage to human life, property and environment caused by the running state of the system in the process of human production below the acceptable level. In another word, safety usually refers to that person is not threatened, in danger, harmed or in loss.

We hope that our study can provide new ideas for clinical treatment under the permission of safety. Therefore, in this chapter we will describe how we address biosecurity and other safety requirements in our project.

Project safety

Our project is dedicated to the study of oncolytic viruses M1, and it was designed and conducted with our professors’ permission and advice.M1 is a strain of Getah-like alphavirus that was isolated from culicine mosquitoes collected on Hainan Island of China [1，2]. Getah virus is transmitted mainly among horses and pigs, and it has not been linked to human illness [3-5]. Also, M1 does not cause apparent disease symptoms in mice or rats, even on administration of doses up to 3 × 107 pfu per mouse or 3 × 108 pfu per rat [1]. M1 was identified as a novel selective killer targeting zinc-finger antiviral protein (ZAP)-deficient cancer cells, and large-scale multicenter pathology study using tissue microarrays reveals that ZAP is commonly deficient in human cancer cells while not in normal cells [6]. Thus M1 kills tumor cells, but activates innate immunity that can eliminate M1 in normal cells which makes it safe to human. Therefore, our research is harmless to the human body and the environment. Though M1 is not included in the White List, we have gained the permission by checking-in. You can click here to see the Check-in Form. All the competent cells (TOP10,JM110,DH5A,BL21) we used are non-pathogenic and safe to human.

Design safety

As a Chinese saying goes, we should nip something harmful in the bud. Foreseeing safety problems and proposing solutions is a necessity for everyone engaged in biological research, and that's what we did. The following table shows the security issues we considered in the design and the corresponding solutions.

problem	Solution
Physical safety (like high pressure steam, sharp instrument, inflammable)	Follow standard operating procedures Wear necessary protective equipment: protective clothing, insulating gloves, masks, etc Do not reinsert or amputate the used needle No open fire in flammable area Cut off the source of hazard in time when combustion occurs
Chemical safety (like toxic chemicals, anesthetics, disinfectant)	Manage and use strictly in accordance with national regulations and standard operating procedures Storage containers should be marked accordingly Wear necessary protective clothing, gloves, masks, etc Avoid prolonged or direct contact Equipped with appropriate instruments for auxiliary use Operate in fume hood or biosafety cabinet
Biosafety (like allergy, pollution waste, microorganisms, bites or scratches)	Use microorganisms and plasmids that are harmless to humans High pressure sterilization of infectious waste Standard operation and personal protection Clean and disinfect strictly before and after the experiment Wash the wound with plenty of water and soap in time and seek medical advice and vaccinate if any bites or scratches injury

Lab safety

Our project is conducted in the biosafety 1/2 laboratory (BSL-2, according with the national standard GB 19489-2008, visiting http://kj.medste.gd.cn/sciedu/ArticleShow.asp?ArticleID=258971 to see detials), and our lab conforms to the regulations on laboratory safety management of Sun Yat_Sen university(see details in http://sbc.sysu.edu.cn/rules/security). The experimental consumables especially the quick-burning, flammable materials are stored in designated locations away from the fire source, and the lab is equipped with effective firefighting facilities. The laboratory and office are strictly separated to prevent unnecessary cross-contamination. Our lab provides biosafety cabinets, autoclave cookers and eye washers, and the waste generated in the experiment was put into trash bags and dustbins with biohazard labels which disposed by relevant personnel which conforming the Do Not Release Policy.

Our team is made up of members from various disciplines, and members in the experimental group have been trained about general lab safety and experimental skills by our guide teacher such as proper use of laboratory equipment and so on. For the safety and health of experimenters in our team, eating and drinking are prohibited in the laboratory, and all the experimenters must wear protective equipment such as gloves and Lab coats. All laboratory work was in accordance with the safety requirements for the laboratory.

Ethics

Laboratory animals are replacements for humans. Therefore, safeguarding animal welfare is not only a humanitarian requirement, but also related to the quality of scientific research. Our project was consistent with the national standard GB/T 35892-2018 (Laboratory animal—Guideline for ethical review of animal welfare, you can see details by visitingwww.gb688.cn/bzgk/gb/newGbInfo?hcno=9BA619057D5C13103622A10FF4BA5D14 ) and approved by the Animal Ethical committe. You can click here to see the Animal Use Form we have finished.

REFERENCE

[1] Hu J, Cai XF, Yan G. (2009) Alphavirus M1 induces apoptosis of malignant glioma cells via downregulation and nucleolar translocation of p21WAF1/CIP1 protein. Cell Cycle 8(20):3328–3339

[2] Wen JS, et al.(2007) Genomic analysis of a Chinese isolate of Getah-like virus and its phylogenetic relationship with other Alphaviruses. Virus Genes 35(3):597–603

[3] Fukunaga Y, Kumanomido T, Kamada M(2000) Getah virus as an equine pathogen. Vet Clin North Am Equine Pract 16(3):605–617

[4] Strauss JH, Strauss EG(1994) The alphaviruses: Gene expression, replication, and evolution. Microbiol Rev 58(3):491–562

[5] Zhai YG, et al. (2008) Complete sequence characterization of isolates of Getah virus (genus Alphavirus, family Togaviridae) from China. J Gen Virol 89(Pt 6):1446–1456