## 3 Results
### Measurement of Cytokine Expression in Spleen Cells  
Similar to the cytokine secretion-promoting effect pattern in the serum described above (Table 7), the expression of spleen cell cytokines treated with the FRB group (0.5 g/kg-b.w., respectively), including IL-1\(\alpha\) (\(p\) < 0.05), IL-2 (\(p\) < 0.05), IL-12 (\(p\) < 0.05), IFN-\(\gamma\) (\(p\) < 0.05), and TNF-\(\alpha\) (\(p\) < 0.01), was also significantly increased (Figure 3a,c,g,h). The FRB group showed higher activity than those of the RB in the expression of cytokines evaluated in this study.  
\begin{table}
\begin{tabular}{c c c c c} \hline \hline (pg/mL) & **Normal** & **CY Control** & **CY + FRB** & **CY + RB** \\ \hline IL-1\(\alpha\) & 58.4 \(\pm\) 7.2 ab & 50.6 \(\pm\) 2.8 b & 62.7 \(\pm\) 5.1 a,* & 52.2 \(\pm\) 3.0 b \\ IL-1\(\beta\) & 9.4 \(\pm\) 0.8 a & 6.2 \(\pm\) 1.3 b & 6.8 \(\pm\) 0.2 b & 7.0 \(\pm\) 0.4 b \\ IL-2 & 38.6 \(\pm\) 3.9 ab & 30.3 \(\pm\) 4.8 b & 39.4 \(\pm\) 1.8 a,* & 37.4 \(\pm\) 2.1 ab \\ IL-4 & 26.5 \(\pm\) 1.4 a & 10.8 \(\pm\) 0.6 c & 13.6 \(\pm\) 1.1 b,** & 13.4 \(\pm\) 0.7 b,** \\ IL-6 & 157.1 \(\pm\) 18.8 a & 96.4 \(\pm\) 11.5 b & 169.1 \(\pm\) 13.5 a,** & 144.6 \(\pm\) 8.8 a,** \\ IL-12 & 259.2 \(\pm\) 13.7 a & 177.5 \(\pm\) 9.0 b & 255.8 \(\pm\) 14.1 a,** & 251.9 \(\pm\) 13.3 a,** \\ (p70) & & & & \\ IFN-\(\gamma\) & 379.5 \(\pm\) 24.6 a & 180.6 \(\pm\) 22.5 c & 351.2 \(\pm\) 22.0 a,** & 222.8 \(\pm\) 9.4 b,** \\ TNF-\(\alpha\) & 12.5 \(\pm\) 1.3 a & 7.7 \(\pm\) 0.8 c & 15.6 \(\pm\) 0.9 b,** & 11.4 \(\pm\) 1.1 a,* \\ \hline \hline \end{tabular}  
* Different letters indicate statistically significant differences between groups using one-way ANOVA followed by [PERSON]’s test of \(p\) < 0.05. The results are expressed as the mean \(\pm\) standard deviation of 5 animals per group. Significantly different from CY Control group (\(p\) < 0.05 and \(*\) \(p\) < 0.01). (CY control: CY only, CY + FRB: CY with FRB 0.5 g/kg-bw, CY + RB: CY with RB 0.5 g/kg-bw, CY with RB 0.5 g/kg-bw).  
\end{table}
Table 7: Profile of cytokines in BALB/c mice after 14 days on the experimental immunology.  
Figure 3: Real time PCR quantitative analysis of cytokine mRNA expression in the spleen. For real-time PCR, we used an SYBR green mix with cytokine gene-specific primers (**(a)**: IL-14\(\alpha\), **(b)**: IN-1\(\beta\), **(c)**: IL-2, **(d)**: IL-4, **(e)**: IN-6, **(f)**: IL-12, **(g)**: IFN-\(\gamma\), **(h)**: TNF-\(\alpha\)). The level of mRNA expression was normalized with GAP-H. Each value is expressed as mean \(\pm\) standard deviation. \({}^{\text{a-c}}\) Different letters indicate statistically significant differences between groups using one-way ANOVA followed by [PERSON]’s test of \(p<0.05\). Significantly different from the CY Control group (\({}^{\text{*}}p<0.05\), \({}^{\text{**}}p<0.01\)). (NC: normal control, CY: negative control (CY only), GS: positive control (CY with Ginseng extract 0.5 kg/kg-bw), FRB: CY with FRB 0.5 g/kg-bw, RB: CY with RB 0.5 g/kg-bw).  
The early response cytokines, TNF-\(\alpha\) and IL-1, are generated in response to microbial challenges. These cytokines amplify, propagate, and coordinate proinflammatory signals, resulting in the synchronized expression of effector molecules that mediate diverse aspects of innate immunity [23]. IL-1 is well known as the innate mediator of T cell immunity. IL-12 is a heterodimeric pro-inflammatory cytokine that regulates T-cell and natural killer-cell responses, induces the production of IFN-\(\gamma\), favors the differentiation of T helper 1 (T\({}_{\rm H}\)1) cells, and is an important link between innate resistance and adaptive immunity [7; 23].