**2.6 Administration method of** β**-glucan**

Dissolve the extract in physiological saline, and for the control group, tumor-seeding group, β-glucan administration group, β-glucan + tumor seeding group, extract the water extract (200 mg/kg) every other day for several weeks. Administered. In the control group, tumor seeding +2 Gy-exposed group and β-glucan + tumor seeding +2 Gy-exposed group, only saline was injected. After 2 weeks of cancer cell seeding, the tumor size was measured weekly. We also measured body weight on exactly the same schedule as tumor size measurements.

Each group consisted of 10 mice. Tumor size was measured using. Formula:

( ) <sup>3</sup> Tumor size cm = 3/4 πA2B/2

A; Minor axis (cm);

B; Longer axis (cm)

At 14 (just before the first X-ray exposure), 1, 3, 5, 7, 9 and 11 days after tumor dissemination, each mouse is fixed with a holder and the caudal vein is covered with an extended female to get about 20 ul of dripping blood. Blood was collected in a blood collection container (Dolamond Co. Ltd.) and diluted with diluent (Nihon Kohden Co. Ltd.). White blood cell counts, including lymphocyte counts and granulocyte counts, were measured using an automated blood cell analyzer (Nihon Kohden Co. Ltd., Celltac-α-MEK-6318). The results obtained were expressed as mean ± standard deviation (S.D.). Student's t-test was applied and results above 0.05 were considered significant.

#### **3. Results**

#### **3.1 Number of peripheral blood cells: leukocytes**

**Figure 1** shows the changes over time of leukocytes in the 4 groups. Compared with the control group, the β-glucan group showed a significant increase in the number of white blood cells. The rate of significant increase throughout the treatment period was 5/8 (β-glucan group). These results suggest that β-glucan decisively increases white blood cell count.

**105**

**Figure 2.**

*New Screening for the Development of Radioprotectors: Radioprotection and Anti-Cancer Effect…*

In the irradiated group, white blood cell counts began to decrease in all experimental groups immediately after irradiation and tended to recover after 3 days. Compared to the 2 Gy group over time, the white blood cell count was always higher in the β-glucan +2 Gy group. When observed in each period, the white blood cell count in the β-glucan +2 Gy group was significantly higher than the 2 Gy single group the day before irradiation (*P < 0.05*). This suggests that an increase in white blood cell count is shown in the un-irradiated state. A significant ratio was observed in the β-glucan +2 Gy group. These results suggest a decrease in 2 Gy-induced

β*-glucan on blood leukocyte counts in mice. There were 10 animals in each experimental group. Data are mean ± standard deviation values. Statistically significantly different (\* P < 0.05) from the control group. Statistically significantly different (\* P < 0.05) from the 2 Gy group. Administration of* β*-glucan was administered for 2 weeks and then treated with radiation. da; days after, hp.; hours post, dp; days post.*

**Figure 2** shows the time course of lymphocyte counts by β-glucan administra-

β*-Glucan on blood lymphocyte counts in mice. There were 10 animals in each experimental group. Data are mean ± standard deviation values. Statistically significantly different (\* P < 0.05) from the control group. Statistically significantly different (\* P < 0.05) from the 2 Gy group. Administration of* β*-glucan was administered for 2 weeks and then treated with radiation. da; days after, hp.; hours post, dp; days post.*

In the non-irradiated group, all treatment groups showed higher lymphocyte counts compared to the control group. Specifically, there was a significant ratio in the β-glucan

decrease in white blood cell count in the β-glucan +2 Gy group.

tion in each non-irradiated and 2 Gy whole body irradiated groups.

**3.2 Number of lymphocytes**

**Figure 1.**

*DOI: http://dx.doi.org/10.5772/intechopen.95016*

*New Screening for the Development of Radioprotectors: Radioprotection and Anti-Cancer Effect… DOI: http://dx.doi.org/10.5772/intechopen.95016*

#### **Figure 1.**

*Translational Research in Cancer*

The X-ray generator used was Philips MG226/4.5. For the pipe voltage, a dose of 200 kV was applied at a rate of 0.35 Gy/min. The total amount of X-rays exposed was 2 Gy. The exposed place of the mouse was fixed to the position of the front part of the left foot (place where cancer cells were seeded) with a holder, and the place not irradiated was covered with a lead container. X-rays were irradiated on the 1 and 3 days. The specific radiation irradiation method in each experimental group is shown below. For changes in blood cells, single irradiation with 2 Gy was performed. However,

In order to obtain reproducible experimental data, mice were inoculated on the

cancer cells were inoculated into the muscle of the left foot of an ICR male mouse.

( ) <sup>3</sup> Tumor size cm = 3/4 πA2B/2

At 14 (just before the first X-ray exposure), 1, 3, 5, 7, 9 and 11 days after tumor dissemination, each mouse is fixed with a holder and the caudal vein is covered with an extended female to get about 20 ul of dripping blood. Blood was collected in a blood collection container (Dolamond Co. Ltd.) and diluted with diluent (Nihon Kohden Co. Ltd.). White blood cell counts, including lymphocyte counts and granulocyte counts, were measured using an automated blood cell analyzer (Nihon Kohden Co. Ltd., Celltac-α-MEK-6318). The results obtained were expressed as mean ± standard deviation (S.D.). Student's t-test was applied and results above

Dissolve the extract in physiological saline, and for the control group, tumor-seeding group, β-glucan administration group, β-glucan + tumor seeding group, extract the water extract (200 mg/kg) every other day for several weeks. Administered. In the control group, tumor seeding +2 Gy-exposed group and β-glucan + tumor seeding +2 Gy-exposed group, only saline was injected. After 2 weeks of cancer cell seeding, the tumor size was measured weekly. We also measured body weight on exactly the same schedule as tumor size measurements. Each group consisted of 10 mice. Tumor size was measured using.

sarcoma 180

in the radiation tumor effect experiment, 2 Gy was divided into 3 divided doses.

15th day after breeding for more than 1 month. Approximately 2 x 106

**2.4 Irradiation device**

**2.5 Tumor inoculation**

Formula:

A; Minor axis (cm); B; Longer axis (cm)

0.05 were considered significant.

**3.1 Number of peripheral blood cells: leukocytes**

that β-glucan decisively increases white blood cell count.

**Figure 1** shows the changes over time of leukocytes in the 4 groups.

Compared with the control group, the β-glucan group showed a significant increase in the number of white blood cells. The rate of significant increase throughout the treatment period was 5/8 (β-glucan group). These results suggest

**2.6 Administration method of** β**-glucan**

**104**

**3. Results**

β*-glucan on blood leukocyte counts in mice. There were 10 animals in each experimental group. Data are mean ± standard deviation values. Statistically significantly different (\* P < 0.05) from the control group. Statistically significantly different (\* P < 0.05) from the 2 Gy group. Administration of* β*-glucan was administered for 2 weeks and then treated with radiation. da; days after, hp.; hours post, dp; days post.*

In the irradiated group, white blood cell counts began to decrease in all experimental groups immediately after irradiation and tended to recover after 3 days. Compared to the 2 Gy group over time, the white blood cell count was always higher in the β-glucan +2 Gy group. When observed in each period, the white blood cell count in the β-glucan +2 Gy group was significantly higher than the 2 Gy single group the day before irradiation (*P < 0.05*). This suggests that an increase in white blood cell count is shown in the un-irradiated state. A significant ratio was observed in the β-glucan +2 Gy group. These results suggest a decrease in 2 Gy-induced decrease in white blood cell count in the β-glucan +2 Gy group.

#### **3.2 Number of lymphocytes**

**Figure 2** shows the time course of lymphocyte counts by β-glucan administration in each non-irradiated and 2 Gy whole body irradiated groups.

In the non-irradiated group, all treatment groups showed higher lymphocyte counts compared to the control group. Specifically, there was a significant ratio in the β-glucan

#### **Figure 2.**

β*-Glucan on blood lymphocyte counts in mice. There were 10 animals in each experimental group. Data are mean ± standard deviation values. Statistically significantly different (\* P < 0.05) from the control group. Statistically significantly different (\* P < 0.05) from the 2 Gy group. Administration of* β*-glucan was administered for 2 weeks and then treated with radiation. da; days after, hp.; hours post, dp; days post.*

#### *Translational Research in Cancer*

group (4/8), and statistically significant differences were observed over many periods (*P < 0.05*). Overall, lymphocytes tended to increase compared to the control group.

This is the same trend seen in the non-irradiated group. Time-dependent changes after irradiation, all experimental groups showed a decrease, and subsequent recovery was observed 3 days after irradiation. The β-glucan +2 Gy group also showed higher lymphocyte counts than the X group. A significant ratio was observed in the β-glucan +2 Gy group (*P < 0.05*). These treatments have been shown to be particularly effective with a recovery period of 3 days after irradiation.

#### **3.3 CD4 + and CD8 +**

**Figures 3** and **4** show the changes in CD4 + and CD8 + after β-glucan administration compared to the control group.

**Figures 3** and **4** shows the increase frequency of CD4 + or CD8 + by β-glucan administration in each non-irradiated and 2 Gy whole body irradiation groups.

The β-glucan +2 Gy group showed a higher increase in CD4 + or CD8 + than the X group.

**Figure 3.**

*Lymphocytes were analyzed for CD4+ in C3H mic. The change of CD4+ cells after control and* β*-glucan administration group. The change of CD4+ cells control and* β*-glucan administration group. Statistically significantly different (\* P < 0.05) from the control group.*

#### **Figure 4.**

*Lymphocytes were analyzed for CD8+ in C3H mic. The change of CD8+ cells after control and* β*-glucan administration group. The change of CD8+ cells control and* β*-glucan administration group. Statistically significantly different (\* P < 0.05) from the control group.*

**107**

*New Screening for the Development of Radioprotectors: Radioprotection and Anti-Cancer Effect…*

Although an effect was observed in the β-glucan administration group, there was no statistically significant difference. The antioxidant activity against peroxy

*Effect of* β*-glucan on the tumor growth in mice inoculated with 4 T1 (high grade) of mouse cancer cell line. Groups of ten mice each were subjected to each treatment. Results represent means ± S.D. \* Statistically* 

**Figure 5** shows the measurement of tumor growth rate by β-glucan administration. The doubling time of the non-irradiated group was not different from the doubling time of the control group with respect to the number of days required to double the tumor size and the ratio of the control group to the 6 Gy group. In the local irradiation group (treatment group), the doubling time of the β-glucan +6 Gy group was 1.4 times longer than that of the control group. The doubling time of the β-glucan +6 Gy group did not exceed the doubling time of the 6 Gy group and tended to be slightly shorter. Compared to the 6 Gy group, it was confirmed that

White blood cells consisting of lymphocytes, granulocytes, and monocytes are deeply involved in immunity. Lymphocytes are roughly classified into T cells and B cells. T cells are associated with cellular immunity, while B cells are associated with humoral immunity through the production of antibodies. T cells are further classified into helper T cells and suppressor T cells, helper T cells play a role in directing and activating B cells, NK cells, killer T cells, cytotoxic T cells, and granulocytes are mediated by blood Phagocytoses vascular walls, bacteria and foreign bodies. Monocytes are transformed into macrophages through morphological changes after moving to tissues, and transmit antigen information to T lymphocytes. In addition, macrophages activate NK and LAK cells. Thus, in white blood cells, lymphocytes distinguish between self and non-self, give instructions, and play a central role in the immune response [15–19]. In this study, in the non-irradiated group, the β-glucan group showed an increase in the number of lymphocytes, suggesting an

*DOI: http://dx.doi.org/10.5772/intechopen.95016*

**3.4 Effect of antioxidant activity (AOAAAPH)**

radicals after treatment was subtle.

*significant (P < 0.05) from the control.*

tumor growth was suppressed.

increase in the overall white blood cell count.

**4. Discussion**

**3.5 Anti-tumor effects**

**Figure 5.**

*New Screening for the Development of Radioprotectors: Radioprotection and Anti-Cancer Effect… DOI: http://dx.doi.org/10.5772/intechopen.95016*

**Figure 5.**

*Translational Research in Cancer*

**3.3 CD4 + and CD8 +**

X group.

**Figure 3.**

tration compared to the control group.

group (4/8), and statistically significant differences were observed over many periods (*P < 0.05*). Overall, lymphocytes tended to increase compared to the control group. This is the same trend seen in the non-irradiated group. Time-dependent changes after irradiation, all experimental groups showed a decrease, and subsequent recovery was observed 3 days after irradiation. The β-glucan +2 Gy group also showed higher lymphocyte counts than the X group. A significant ratio was observed in the β-glucan +2 Gy group (*P < 0.05*). These treatments have been shown

to be particularly effective with a recovery period of 3 days after irradiation.

**Figures 3** and **4** show the changes in CD4 + and CD8 + after β-glucan adminis-

**Figures 3** and **4** shows the increase frequency of CD4 + or CD8 + by β-glucan administration in each non-irradiated and 2 Gy whole body irradiation groups.

*Lymphocytes were analyzed for CD8+ in C3H mic. The change of CD8+ cells after control and* β*-glucan administration group. The change of CD8+ cells control and* β*-glucan administration group. Statistically* 

*Lymphocytes were analyzed for CD4+ in C3H mic. The change of CD4+ cells after control and* β*-glucan administration group. The change of CD4+ cells control and* β*-glucan administration group. Statistically* 

*significantly different (\* P < 0.05) from the control group.*

*significantly different (\* P < 0.05) from the control group.*

The β-glucan +2 Gy group showed a higher increase in CD4 + or CD8 + than the

**106**

**Figure 4.**

*Effect of* β*-glucan on the tumor growth in mice inoculated with 4 T1 (high grade) of mouse cancer cell line. Groups of ten mice each were subjected to each treatment. Results represent means ± S.D. \* Statistically significant (P < 0.05) from the control.*
