Paper Anonymously posted by Chinese Whistleblower, Dr. Li-Meng Yan on 1/25/2020 Proving that the PLA-owned Zhoushan bat virus was the closest relative to 2019-nCoV before RaTG13 mysteriously appeared out of nowhere - "Brief summary of evidence of lab-made 2019 nCoV"

Brief summary of evidence of lab-made 2019 nCoV

From 2017-2019, a series academic papers about a novel zoonotic coronavirus isolated from bats in Zhoushan, Zhejiang (in short, ZS bat-CoV) were published in English (1) and Chinese

(1. post-graduate thesis: Preliminary study about bat-virus in South-east coastland http://le.cnki.net/kmobile/Master/detail/SYJT_PHAM/1017235765.nh)

(2. Molecular identification and analyse of bat-coronavirus in Zhoushan area http://kns.cnki.net/kcms/detail/detail.aspx?filename=JSCY201901004&dbcode=CJFQ &dbname=CJFDTEMP&v=).

The main investigators were from Chinese military institution (The Third Military Medical University and Military medical institute of Nanjing Command) (Pic. 1-3).

It means the novel ZS bat-CoV was owned by Chinese military labs only. In the papers above, they have clarified that ZS bat-CoV has potential cross-species transmission, including human, also emphasized that it is significant for public health. The full genome sequence of certain strains of ZS bat-CoV were released in Genbank of NIH (accession numbers MG772844 through MG772934), provided by Nanjing command (Pic. 4-5).

11 Jan 2020, China CDC released the full genome sequence of 2019 nCoV, and it was upload to GenBank of NIH on 12 Jan (Wuhan seafood market pneumonia virus isolate Wuhan-Hu-1, accession number MN908947.1). 14 Jan, MN908947.1 was replaced by China CDC without announced (accession number MN908947.2). 17 Jan, MN908947.2 was replaced by them for unknown reasons (accession number MN908947.3) *(pic. 6-7).

The blast results based on full genome of MN908947.2 and MN908947.3 shown that ZS bat-CoV is the most closed known coronavirus to 2019 nCoV via online blast toolNIH (per Identity is 88.65-89.12%, MG772933.1 and MG772934.1) (pic. 8). According to analysis of virus evolution tree, ZS bat-CoV is the most closed relative to 2019 nCoV (https://nextstrain.org/groups/blab/sars-like-cov) (pic. 9). To note, when blast MN908947.1 at that time, there were some error in the sequence which affected the results (no picture recorded).

Further blast results for the most critical proteins and segments between 2019 nCoV and all the other known coronavirus in Genbank, still show the most significant similarity to ZS bat-CoV, which is much higher than compared with any others. The similarity is Spike protein 80.32-81.00% (S protein) (pic. 10-12) and Nucleocapsid protein 94.03% (pic. 13-15) to two strains of ZS bat-CoV (MG772933.1 and MG772934.1), while Envelop protein 100% (E protein) (pic. 16-18), ORF8 segment 94.21% (pic. 19-20), and Membrane glycoprotein 98.65% (pic. 21-22) to MG772933.1, as well as RdRp gene 95.75% (pic. 23-24) to MG772934.1.

As mentioned in Nanjing military paper(1), from homology analyses of different ORFs, ORF8 fragments in ZS bat-CoV showed the lowest homology, presenting only 60% identity with its closest relatives (while 94.21% to 2019 nCoV) (pic. 25). As well, analysis of the RNA-dependent RNA polymerase (RdRp) gene showed that the genomic sequences of bat CoV samples obtained from different parts of the world shared 80–90% identity among themselves and exhibited 87–92% identity with the SARS-CoVs extracted from human or civet sources (while 95.75% to 2019 nCoV).

Compared with high mutant S protein, E protein is more conserved (2). However, undergoing a natural evolution, the possibility of 100% identical E protein between the cross-species 2019 nCoV and ZS bat-CoV is almost impossible. It was simply confirmed by using another online tool-Cluster Omega, which shows that the E protein of one strain of ZS bat-CoV (AVP78033.1, belong to MG772933.1) is more identical (100%) to 2019 nCoV, rather than another ZS bat-CoV (AVP78044.1, belong to MG772934.1) (pic. 26).

21 Jan, the first paper studied 2019 nCoV is published in a journal under Chinese Academy of Sciences, from Beijing Institute of Pharmacology and Toxicology (belonging to military) and Chinese Academy of Sciences (3). In that paper, they mentioned that the natural host of 2019 nCoV may be certain bat (not directly mentioned ZS bat-CoV). In their official Chinese introduction for this paper, they stressed the key point of their finding as: “To be surprised”, compared with SARS CoV, 4 of 5 amino acid changed in the receptor binding domain (RBD) of S protein in 2019 nCoV, but still maintains the core structure to support strong interaction with human ACE2 molecules “in a very perfect way”. It means 2019 nCoV could infect human respiratory epithelial cells in the same way as what SARS CoV did (pic. 27).

To noted, 23 Jan, another paper with similar content was upload to BioRxiv, which is from famous bat and SARS CoV investigator Zheng-li Shi’s team (4).

There are also a lot of official news and poor-quality academic articles from end of last Dec show that no evidence about wild animals in Huanan seafood market as intermediate host for 2019 nCoV (which can be explained in a detail way later). Hence, one hypothesis of lab-made 2019 nCoV is recombined with SARS RBD of S protein (to human ACE2 gene), based on ZS bat-CoV (esp. MG772933.1), going through in vitro and in vivo adaptation and amplification in a limited range in the lab, generated an ideal strain (2019 nCoV) with effective RBD, while the other comparable conserved sequence did not change much, or even without any change (E protein). Since stock virus kept in culture media at -80 ℃, slowly thaw it on ice could help the virus released in the environment better.

According to Mr. Miles Guo, thymosin is out of stock in commercial market in China. Thymosin is potential therapeutic in the treatment of severe SARS and MERS (5).

PS. More evidence from official media and government response along the time in 2019 nCoV crisis is not written in this brief report.

See the actual report for pictures.

/r/China_Flu Thread Link - s3-ap-northeast-1.amazonaws.com