# In Vitro Genotoxicity Assessment from the _Glycyrrhiza_ New Variety Extract
## 4 Discussion
\begin{table}
\begin{tabular}{c c c c c c c c} \hline \hline
**Dose** & **Animals** & \multicolumn{3}{c}{**Body Weight (g) at the Time of**} & \multicolumn{2}{c}{**MNPCE/4000 PCE**} & \multicolumn{1}{c}{**PCE:RBC Ratio**} & \multicolumn{1}{c}{**(\% Control)**} \\ **(mg/kg/Day)** & **per Dose** & **Day 0** & **Day 1** & **Sacrific** & **(Mean \(\pm\) SD)** & **(Mean \(\pm\) SD)** & **(Mean \(\pm\) SD)** & \\ \hline
0 & 6 & 35.34 \(\pm\) 0.96 & 35.34 \(\pm\) 1.61 & 35.43 \(\pm\) 1.40 & 3.83 \(\pm\) 1.47 & 0.52 \(\pm\) 0.02 & 100 \\
1250 & 6 & 35.09 \(\pm\) 1.21 & 34.35 \(\pm\) 1.32 & 33.78 \(\pm\) 2.18 & 3.33 \(\pm\) 2.16 & 0.52 \(\pm\) 0.03 & 100 \\
2500 & 6 & 34.98 \(\pm\) 1.16 & 34.93 \(\pm\) 1.25 & 34.81 \(\pm\) 1.12 & 4.17 \(\pm\) 1.47 & 0.52 \(\pm\) 0.03 & 100 \\
5000 & 6 & 34.75 \(\pm\) 1.36 & 34.99 \(\pm\) 1.41 & 35.25 \(\pm\) 1.61 & 4.00 \(\pm\) 1.79 & 0.53 \(\pm\) 0.02 & 102 \\
**Positive Control** & & & & & & & \\ CPA 70 & 6 & 34.63 \(\pm\) 1.23 & 35.08 \(\pm\) 1.21 & 35.26 \(\pm\) 1.29 & 62.17 \(\pm\) 8.80 ** & 0.41 \(\pm\) 0.03 ** & 78 \\ \hline \hline \end{tabular}  
* * Significantly different from the negative control group at \(p<0.01\). Vehicle, Sterile distilled water for injection. Vehicle and Test article were orally administered to mice for two consecutive days. CPA was intraperitoneally administered to mice once on the day of the 2 nd admin. Mice were euthanized approximately 24 h after the final administration. Bone marrow smears were prepared approximately 24 h after the final administration. MNPCE, Micronucleated polychromatic erythrocyte; PCE, Polychromatic erythrocyte; RBC, Red blood cells (polychromatic erythrocyte + normochromatic erythrocyte); CPA, Cyclophosphamide monohydrate (positive control article).  
\end{table} 
Table 3: In vivo bone marrow micronucle test of WG extract. mutations or point mutations, including the addition, deletion, or substitution of one or more DNA base pairs [35]. The _S. typhimurium_ strains used in the original assay have a mutation in the histidine synthesis gene and are auxotrophic to histidine. When the bacteria are exposed to a potential mutagen and plated in histidine-limited media, colony forming is developed because bacteria mutated and reverted to being prototrophic to histidine. A similar process is followed with respect to the tryptophan auxotroph strains of _E. coli_. Accordingly, counting colony units is a means of assessing mutagenic potential. Test substances are evaluated with or without S9 mix to investigate promutagens and direct mutagen activity [36,37]. Compared with a negative control group, we observed no cytotoxicity and an increase of the number of bacterial revertants in _S. typhimurium_ (TA100, TA1535, TA98, and TA1537) and _E. coli_ (WP2 _uvr_A) up to 5000 \(\upmu\)g/plate, regardless of the presence of S9 mix (Table 1), suggesting that WG extract didn't induce the base pair substitution mutations (TA100, TA1535 and WP2 _uvr_A) and frameshift mutations (TA98 and TA1537) under in vitro conditions of this study. Also, the potential mutagenic properties of WG extract were evaluated using a chromosome aberration test. Chromosome aberrations correlate with genetic disorders like cancer, which causes structural and numerical chromosome aberrations [38]. Abnormal replication or repair of DNA lesions can directly or indirectly induce double-stranded DNA damage, which leads to breakage of the double-strand structural. This is the primary cause of structural chromosome aberrations [39]. The disruption of cell cycle checkpoints or the inhibition of topoisomerase damage to DNA, which is the cause of the numerical chromosome aberrations [40]. A chromosome aberration test is able to identify structural and numerical chromosome aberrations using cultured mammalian cells, and is accordingly used to assess the genotoxicity of candidate compounds. CHL cells are commonly used in chromosome aberration tests due to their sensitivity to mutagens and their low chromosome number, which facilitates scoring [41].