The lymphocytes exposed to BE <i>in vivo</i> underwent increased apoptosis upon subsequent culture <i>in vitro</i>.

<p>Murine hepatitis was induced by an intravenous injection of Con A at a dose of 15 mg/kg. BE (100 mg/kg) was administrated intraperitoneally immediately after Con A injection. The liver MNCs and splenocytes were prepared at 8 h after Con A injection. After 2 hours of in-vitro culture, the adherent macrophages were removed and the nonadherent cells were further cultured <i>in vitro</i> for 12 and 24 h. (A) The percentages of apoptosis in the <i>in vitro</i> cultured nonadherent cells at each time point were determined by flow cytometry using FITC-annexin V/PI staining. (B) After <i>in vitro</i> culture for 24 h, the absolute numbers of apoptotic cells in liver MNCs and splenocytes, as well as in CD3⁺ T cell and CD19⁺ B cell subsets, were determined by cell counting and flow cytometry using PE-anti-CD3/FITC-annexin V staining and PE-anti-CD19/FITC-annexin V staining, respectively. Data are mean ± SEM of 6–8 mice/group. *<i>P</i><0.05 versus Con A only control, ^#<i>P</i><0.05 versus Non-treatment control.</p

BE selectively induces apoptosis of LPS-activated CD19⁺ B cells through mitochondrial pathway.

<p>CD19⁺ B cells were isolated from murine splenocytes using Miltenyi MACS Purification and incubated with indicated concentrations of BE for 24 h in the absence or presence of 10 µg/ml of LPS. (A, B) The percentages of cells with low ΔΨm were analyzed by flow cytometry using JC-1 staining. (C, D) The percentages of active caspase-3+ cells were analyzed by flow cytometry using FITC-anti-active caspase-3 mAb staining. A and C demonstrate representative experiment of three assays. B and D show mean ± SEM of three independent experiments. (E) The release of cytochrome c (Cyto-c) from mitochondria in B cells after BE treatment in the presence of LPS was examined by Western blotting. (F) Protein levels of Bcl-2 and Bax in B cells after BE treatment in the presence of LPS were examined by Western blotting. The results shown in E and F are representative of three experiments. (G) The activities of caspase-3, 8, 9 in B cells after BE treatment in the presence of LPS was measured using colorimetric assay. Each column represents the mean ± SEM of 3 experimental values. *<i>P</i><0.05 versus untreated controls.</p

BE preferentially induces apoptosis in mitogen-activated splennocytes.

<p>Murine splenocytes were treated with indicated concentrations of BE for 24 h in the absence or presence of 5 µg/ml of Con A, and the percentages of apoptosis were detected using Annexin V/PI staining. (A) is a representative of three independent assays. (B) represents mean ± SEM of three independent experiments. *<i>P</i><0.05 versus untreated controls.</p

BE protected mice against Con A-induced hepatitis and increased the incidences of apoptosis in liver-infiltrating MNCs and splenocytes, as well as in CD3⁺ and CD19⁺ splenocytes.

<p>Murine hepatitis was induced by an intravenous injection of Con A at a dose of 15 mg/kg. BE (100 mg/kg) was administrated intraperitoneally immediately after Con A injection. Blood samples were collected at 0, 8, and 24 h after Con A injection. Livers and spleens were collected 24 h after Con A injection. (A) Serum ALT levels at indicated time points after Con A injection. (B) Serum levels of IFN-γ and TNF-α at indicated time points after Con A injection. (C) Photomicrographs of representative H&E stained liver sections (×200). (D) Cell numbers of MNCs, CD3⁺ T cell and CD19⁺ B cells in livers at 24 h after Con A injection. (E) Percentages of apoptosis in liver MNCs detected by flow cytometry using FITC-annexin V/PI staining. (F and G) Flow cytometry analysis of apoptosis in splenic T and B cells using PE-anti-CD3/staining and PE-anti-CD19/FITC-annexin V staining, respectively. F shows representative results of 10 mice in each experimental group. Data in A, B D, E, and G are mean ± SEM of 10 mice/group. *<i>P</i><0.05 versus Con A only control, ^#<i>P</i><0.05 versus Non-treatment control.</p

BE selectively induces apoptosis in activated lymphocytes.

<p>(A) Different types of immune cells as indicated were incubated with 10 µM of BE for 24 h in the absence or presence of activator, and the percentages of apoptosis were detected using Annexin V/PI staining. The increase in the percentages of apoptosis in each type of cells upon BE exposure was calculated. Following activators were used: 5 µg/ml of Con A for splenocytes and CD3⁺ splenocytes; 500 ng/ml of LPS for CD19⁺ splenocytes, BM-derived DCs, peritoneal macrophages, and RAW264.7 cells; 25 ng/mL of PMA and 1 µM of ionomycin for Jurkat T cells. Data are mean ± SEM of three independent experiments. *<i>P</i><0.05 versus BE only control. (B) CD3⁺ splenocytes, CD19⁺ splenocytes, and Jurkat T cells were treated with indicated concentrations of BE for 24 h in the absence or presence of respective activator, and the percentages of apoptosis were detected using Annexin V/PI staining. Data are mean ± SEM of three independent experiments. *<i>P</i><0.05 versus untreated controls.</p

Investigation of HIFU-induced anti-tumor immunity in a murine tumor model-2

<p><b>Copyright information:</b></p><p>Taken from "Investigation of HIFU-induced anti-tumor immunity in a murine tumor model"</p><p>http://www.translational-medicine.com/content/5/1/34</p><p>Journal of Translational Medicine 2007;5():34-34.</p><p>Published online 11 Jul 2007</p><p>PMCID:PMC1939831.</p><p></p>llected 1–2 days after HIFU treatment. 6-μm cryostat sections were cut and stained with anti-CD11c Abs. Then the antibody was visualized using the Anti-Hamster Ig HRP detection kit. The sections were counterstained with hematoxylin

Investigation of HIFU-induced anti-tumor immunity in a murine tumor model-0

<p><b>Copyright information:</b></p><p>Taken from "Investigation of HIFU-induced anti-tumor immunity in a murine tumor model"</p><p>http://www.translational-medicine.com/content/5/1/34</p><p>Journal of Translational Medicine 2007;5():34-34.</p><p>Published online 11 Jul 2007</p><p>PMCID:PMC1939831.</p><p></p>cavitation detection (PCD) signals, and (C) images of tumor tissue injury produced by different HIFU treatment

Investigation of HIFU-induced anti-tumor immunity in a murine tumor model-3

<p><b>Copyright information:</b></p><p>Taken from "Investigation of HIFU-induced anti-tumor immunity in a murine tumor model"</p><p>http://www.translational-medicine.com/content/5/1/34</p><p>Journal of Translational Medicine 2007;5():34-34.</p><p>Published online 11 Jul 2007</p><p>PMCID:PMC1939831.</p><p></p>FSE-labeled immature bone marrow-derived DCs were injected into tumor 1 day after HIFU treatment. The (A) total cell number, (B) total number of DC(CD11ccells), and (C) migrating DC (CFSECD11ccells) recovered in DLN on day 2 were determined by flow cytometry. Data points represent the mean ± SD for each group (n = 8). *P < 0.05 versus DC injection only control group. (D) Representative histogram illustrating the population of CFSECD11ccells within the DLN of mice subjected to different treatments

Investigation of HIFU-induced anti-tumor immunity in a murine tumor model-4

<p><b>Copyright information:</b></p><p>Taken from "Investigation of HIFU-induced anti-tumor immunity in a murine tumor model"</p><p>http://www.translational-medicine.com/content/5/1/34</p><p>Journal of Translational Medicine 2007;5():34-34.</p><p>Published online 11 Jul 2007</p><p>PMCID:PMC1939831.</p><p></p>ors, and induced tumor-specific CTL response (C), and IFN-γ-secreting cells (D) in the spleens of treated mice. C57BL/6 mice were inoculated s.c. on right hind leg with 5 × 10MC38 tumor cells and treated with different HIFU on day 9 of tumor inoculation. Mice were challenged with 1 × 10MC38 tumor cells by s.c. inoculation on the left hind leg one day after HIFU treatment. Both primary and challenged tumor growth were monitored daily. Splenocytes obtained from control and treated mice 10 days after HIFU treatment were re-stimulated with mytomicin-pretreated MC-38 or EL4 tumor cells and CTL and ELISPOTS assays were performed. Results were expressed as mean value ± SD for each group (n = 8). *P < 0.05; **P < 0.001 versus non-treatment control. This experiment is representative of three experiments with consistent results

Investigation of HIFU-induced anti-tumor immunity in a murine tumor model-1

<p><b>Copyright information:</b></p><p>Taken from "Investigation of HIFU-induced anti-tumor immunity in a murine tumor model"</p><p>http://www.translational-medicine.com/content/5/1/34</p><p>Journal of Translational Medicine 2007;5():34-34.</p><p>Published online 11 Jul 2007</p><p>PMCID:PMC1939831.</p><p></p>ing. Bright hyperechoic spots generated by HIFU indicate regions of cavitation