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The areas of islets staining positively (insulin and glucagon respectively) and the whole islet were assessed by IPP6.0. And then ��-cell mass and ��-cell mass were respectively calculated (��-cell mass or ��-cell mass=area of insulin or glucagon staining positively/area of the whole islets). Three islets were randomly selective for analysis for each animal (n=5 for each group). Proliferation of islet cells was assessed by immunohistochemical staining for proliferating cell nuclear antigen (PCNA) using PCNA antibodies (sc-7907, 1:100; Santa Cruz, Texas, USA). Slides were incubated with primary antibodies for 1h at room temperature. After washing, secondary antibodies (1:500, biotin-conjugated goat anti-rabbit IgG; Boster, Wuhan, China) were applied for 30 min at room temperature. The number SB203580 price of PCNA-positive cells per islet section was counted. Three islets were randomly selective for analysis for each animal (n=5 for each group). To evaluate the expression of NADPH oxidase P22phox subunit in islet sections, staining was carried out with P22phox antibodies (sc-20781, 1:100; Santa Cruz, Texas, USA). Slides were incubated with the primary antibodies this website for 1 h at room temperature. After washing, secondary antibodies (1:500, biotin-conjugated goat anti-rabbit IgG; Boster, Wuhan, China) were applied for 30 min at room temperature. The average gray-scale intensities of cells staining positively were measured by IPP 6.0. Three islets were randomly selective for analysis for each animal (n=5 for each group). Statistical analysis Data are shown as mean ��SD. One-way ANOVA and LSD-test were performed to assess differences between groups of the female rats. Two-way ANOVA analysis was performed to make sense of the interacted effect of sexes and HRP for the MSG group (male and female) and the MSG-HRP group (male and female). A value of P of each group are shown in Table 1. Body weights showed no statistical difference between the Con group and MSG groups. Compared with the Con group, MSG rats had reduced body length. Therefore, the Lee��s index, reflecting the extent of obesity, was increased in MSG rats compared with the Con group. No statistical significant difference was observed in body Alizarin weight, length and Lee��s index between the MSG and MSG-HRP groups. Table 1 Physiological parameters of the animals. HRP increases insulin sensitivity and ameliorates the glucose tolerance status The response of blood glucose to OGTT at week 12 of the experimental period is shown in Figure 1A, 1B. The AUC of blood glucose concentration was higher in the MSG group compared with the Con group, although the difference was not statistically significant. The MSG-HRP group displayed significantly lower AUC of blood glucose concentration than the MSG group (P