## 3 Results
### Effects of NEM on Inflammation, Apoptosis, and Cell Proliferation In Vivo
a \(p\) < 0.01 as compared with sham by THSD; b \(p\) < 0.01 as compared with OA by THSD; c \(p\) < 0.01 as compared with sham by DT3; d \(p\) < 0.01 as compared with OA by DT3. Figure 8: Histopathological analysis in femoral and tibial articular cartilage (AC). Representative safranin O (SO)-stained histological images of the femoral and tibial AC with SM. Long arrows indicated the thicknesses of tibia or femur AC. And short arrows indicate the clone formation in the articular surfaces. GP = Growth plate; MN = Meniscus; CL = Crucial ligament. Scale bars = 160 \(\upmu\)m. \begin{table}
\begin{tabular}{l c c c c c} \hline \hline \multicolumn{1}{c}{**Items**} & \multicolumn{4}{c}{**Rankin Scores**} \\ \cline{2-6}
**Groups** & **Surface** & **Cellularity** & **Clone** & **Stain intensity** & **Total \({}^{*}\)** \\ \hline Diclofenac sodium & & & & & \\
2 mg/kg & 1.70 \(\pm\) 0.48 \({}^{\text{ac}}\) & 1.70 \(\pm\) 0.48 \({}^{\text{ac}}\) & 1.70 \(\pm\) 0.48 \({}^{\text{ac}}\) & 1.70 \(\pm\) 0.48 \({}^{\text{ac}}\) & 6.80 \(\pm\) 1.14 \({}^{\text{ac}}\) \\ NEM & & & & & \\
100 mg/kg & 1.20 \(\pm\) 0.42 \({}^{\text{ac}}\) & 1.00 \(\pm\) 0.00 \({}^{\text{f}}\) & 0.90 \(\pm\) 0.32 \({}^{\text{af}}\) & 1.30 \(\pm\) 0.67 \({}^{\text{bc}}\) & 4.40 \(\pm\) 0.52 \({}^{\text{ac}}\) \\
50 mg/kg & 1.70 \(\pm\) 0.48 \({}^{\text{ac}}\) & 1.60 \(\pm\) 0.52 \({}^{\text{af}}\) & 1.80 \(\pm\) 0.42 \({}^{\text{af}}\) & 1.60 \(\pm\) 0.52 \({}^{\text{ac}}\) & 6.70 \(\pm\) 0.67 \({}^{\text{ac}}\) \\
25 mg/kg & 2.10 \(\pm\) 0.57 \({}^{\text{a}}\) & 2.20 \(\pm\) 0.63 \({}^{\text{a}}\) & 2.20 \(\pm\) 0.63 \({}^{\text{a}}\) & 1.90 \(\pm\) 0.57 \({}^{\text{ad}}\) & 8.40 \(\pm\) 1.07 \({}^{\text{ac}}\) \\ \hline \hline \end{tabular}
\end{table} 
Table 3: _Cont._Figure 9: Immunohistochemistryistochemistry for inflammation in vivo. (**A**) Representative immunohistochemistry-chemistrical images of TNF\(\alpha\) and COX-2 cells on the femoral and tibial articular cartilage (AC) with synovial membrane (SM). Scale bars = 80 μm. (**B**) The proportion of TNF\(\alpha\)(+) cells was calculated as a ratio of positive cells to total cells. Values are expressed as means \(\pm\) SD in 10 rats. a \(p<0.01\) as compared with sham by THSD; b \(p<0.01\) and c \(p<0.05\) as compared with OA by THSD; d \(p<0.01\) as compared with sham by DT3; e \(p<0.01\) as compared with sham by DT3. (**C**) The proportion of COX-2(+) cells was calculated as a ratio of positive cells to total cells. Values are expressed as means \(\pm\) SD in 10 rats. a \(p<0.01\) and b \(p<0.05\) as compared with sham by THSD; c \(p<0.01\) as compared with OA by THSD; d \(p<0.01\) and e \(p<0.05\) as compared with sham by DT3; \(tp<0.01\) as compared with the OA by DT3. Figure 10: Immunohistochemistryistochemistry for apoptosis and cell proliferation in vivo. (**A**) Representative immunohistochemical images of the cleaved poly(ADP-ribose) polymerase (PARP)- and bromodeoxyuridine (BrdU)-immunoreactive cells on the femoral and tibial articular cartilage (AC) with SM. Scale bars = 100 \(\upmu\)m. (**B**) The proportion of PARP cells was calculated as a ratio of positive cells to total cells. Values are expressed as means \(\pm\) SD in 10 rats. a \(p\) < 0.01 as compared with sham by THSD; b \(p\) < 0.01 as compared with OA by THSD; c \(p\) < 0.01 as compared with sham by DT3; d \(p\) < 0.01 as compared with the OA by DT3. (**C**) The proportion of BrdU cells was calculated as a ratio of positive cells to total cells. Values are expressed as means \(\pm\) SD in 10 rats. a \(p\) < 0.01 as compared with sham by THSD; b \(p\) < 0.01 as compared with OA by THSD; c \(p\) < 0.01 and d \(p\) < 0.05 as compared with sham by DT3; e \(p\) < 0.01 as compared with the OA by DT3. The immune response to BrdU showed differences in the cartilage and synovium in the OA compared with in the sham group. BrdU-immunopositive cells were decreased in the two types of AC of the OA group but were significantly increased in a dose-dependent manner in the NEM group, excluding DS administration.