Proapoptotic effect of nonthermal pulsed ultrasound on prostate cancer cells in a nude mouse model.

BACKGROUND: Ultrasound (US) can induce cell injury, and we have previously reported that adjusting the pulse repetition frequency (PRF) of ultrasound output can induce prostate cancer cell destruction without causing a rise in the temperature of the irradiated area. In this study, we examined the mechanism of nonthermal ultrasound cell destruction, which was not fully clarified in our previous reports. METHODS: In vitro, we evaluated postirradiation cells immediately after treatment and examined membrane disruption by proliferation assay, LDH assay, and apoptosis assay. In vivo, we injected mice with human LNCaP and PC-3 prostate cancer cells and evaluated the therapeutic effects of US irradiation by H-E staining and immunostaining. RESULTS: Proliferation assays showed inhibition at 3 h postirradiation independently of PRF and cell line (p < 0.05). Quantitative assessment of apoptosis/necrosis by flow cytometry showed widely varying results depending on cell type. LNCaP showed an increase in late apoptosis at 0 h independent of PRF (p < 0.05), while PC-3 showed no significant difference at 0 h. The LDH assay showed an increase in LDH independent of PRF in LNCaP (p < 0.05 respectively), but no significant difference in PC-3. In vivo, tumor volume was compared and a significant reduction was observed at 10 Hz for LNCaP (p < 0.05) and 100 Hz for PC-3 (p < 0.001) at 3 weeks after the start of irradiation. The excised tumors were evaluated with Ki-67, Caspase-3, and CD-31 and showed a significant treatment effect independent of cell type and PRF (p < 0.001 respectively). CONCLUSION: Examining the mechanism behind the therapeutic effect of US irradiation revealed that the main effect was achieved by apoptosis induction rather than necrosis.

Combined Treatment with Ultrasound and Immune Checkpoint Inhibitors for Prostate Cancer.

Background: Ultrasound (US) is mostly used for diagnostic purpose but could be used for cancer treatments with a US intensity or frequency fitted to such a purpose. Prostate cancer (PC) has the highest prevalence in the urological field, but indications for immune checkpoint inhibitors (ICIs) for PC are limited to very few cases. In this study, we compared the antitumor effect of US irradiation alone with the combined use of US and ICIs in vitro and in vivo. Methods: PC cell line TRAMP-C2 cells were used in our experiments. TRAMP-C2 cells were irradiated with US with pulse repeated frequencies (PRF) of 1, 10, and 100 Hz. Cell proliferation was evaluated by MTS assay and apoptotic cells were analyzed using flow cytometry. To verify the antitumor effect of US irradiation on PC in vivo, we conducted animal experiments using mice. TRAMP-C2-bearing mice were irradiated with US with PRF of 10 and 100 Hz. Three weeks after the start of US irradiation, anti-PD-1 antibody was administered to the mice. Finally, mice were sacrificed and tumors were collected. Immunohistochemical (IHC) analyses were assessed for cleaved caspase-3 and CD3 in tumor cell extracts. Results: Cell proliferation assays showed that 1 and 10 Hz US significantly inhibited cell survival (p < 0.0001). In addition, US irradiation induced apoptosis at 1, 10, and 100 Hz (p = 0.0129, p = 0.0150, and p = 0.0017, respectively). In animal experiments, a significant tumor growth inhibitory effect was observed at 10 and 100 Hz, and 100 Hz + ICIs (p < 0.05, respectively). Hematoxylin−eosin (H−E) staining showed a significant increase in the necrotic area of the tumor at 100 Hz and 100 Hz + ICIs (p < 0.05, respectively). In addition, under IHC staining the expression level of cleaved caspase-3 and the number of CD3-positive cells increased at 100 Hz (p < 0.05, respectively). Conclusion: US irradiation induced apoptosis in cells and reduced cell viability. In vivo tumor growth was suppressed by combined treatment with US irradiation and ICIs. Further research on immune system activation will lead to less invasive and more efficient treatments for PC.

[SURGEON QUESTIONNAIRE FOR ESTABLISHING A SURGEON EDUCATION SYSTEM FOR TRANSURETHRAL LASER RESECTION OF THE PROSTATE (HoLEP)].

(Introduction)HoLEP's role in the surgical management of benign prostatic hyperplasia (BPH) is steadily growing. In this study, a questionnaire containing questions about perioperative management was submitted to HoLEP surgeons to help establish standard surgical training procedures. (Methods)We sent a comprehensive 17 questionnaires on HoLEP procedures to 18 surgeons. The questionnaire asked, "Which method are you using, the 1-LOBE or 3-LOBE method?", "What educational methods are being used for surgeons?", "How long is the catheter insertion period after HoLEP?", and "What is the most difficult problem encountered in surgical HoLEP education and what aspect of training is the most emphasized?" (Results)Sixteen (88.9%) surgeons answered these questionnaires. Five surgeons reported using the one lobe method, five surgeons reported using the three lobe method, and four surgeons answered that it depends on the case. Regarding educational methods, the main answer was that it is important to evaluate pre-HoLEP imaging tests such as MRI and cystoscopy and to simulate surgery for education. Regarding the postoperative catheter insertion period, 1 day: 1 surgeon, 2 days: 9 surgeons, 3 days: 3 surgeons, 4 days or more: 1 surgeon. The most important thing reported for surgical education was to help beginners understand the characteristics of lasers, including direction, distance to prostate tissue, and adenoma removal. (Conclusions)The surgeons' responses clearly indicated some differences in practices between institutions. More detailed data from these results will provide a step towards designing standardized surgical and educational protocols for HoLEP.

Can ultrasound irradiation be a therapeutic option for prostate cancer?

BACKGROUND: Focal therapies for prostate cancer (PC) can reduce adverse events and do not lead to androgen-independent progression. Ultrasound could be used for cancer treatments if the repetition frequency is fitted to the purpose. We investigated the possible therapeutic effect of ultrasound irradiation on PC cells. MATERIALS AND METHODS: We irradiated two PC cell lines, androgen-dependent LNCaP and -independent PC-3 with ultrasound (3.0 W/cm2 , 3 MHz, irradiation time rate: 20%) for 2 minutes for 1 day or 3 consecutive days at a repetition frequency of 1, 10, or 100 Hz in vitro. Cell proliferation and apoptosis were determined after irradiation. RESULTS: Cell proliferation of PC-3 was significantly inhibited after 1 day (P < .0001) and 3 days (P < .0001) of 10 Hz ultrasound irradiation, and that of LNCaP after 1 day (P < .0001) and 3 days (P < .0001) of irradiation. LNCaP was more sensitive to ultrasound at both lower and higher cell density but PC-3 was only sensitive at a lower cell density (P < .01). Irradiation with 10 Hz ultrasound-induced significantly more PC-3 apoptotic cells than control (1 day, P = .0137; 3 days, P = .0386) rather than irradiation with 1 Hz. Apoptosis via caspase-3 was induced at 10 Hz in 1-day (P < .05) irradiation in both cell lines. CONCLUSIONS: Ultrasound irradiation with even 1 day of 10 Hz significantly inhibited cell proliferation in both LNCaP and PC-3, especially by the remarkable induction of apoptosis in vitro. Our study indicated that ultrasound irradiation can be a therapeutic option for PC and further studies in vivo will be undertaken.

[Diagnostic value of contrast-enhanced ultrasonography in preoperative T-staging of gastric cancer].

OBJECTIVE: To investigate the diagnostic value of contrast-enhanced ultrasonography (CEUS) in preoperative T-staging of gastric cancer. METHODS: A sulfur hexafluoride-filled microbubble ultrasound contrast agent and a continuous real-time imaging technique of contrast pulse sequencing were used. Normal gastric wall was examined by CEUS in 8 healthy volunteers and the results were compared with the findings on multislice computed tomography. Sixty-two patients with gastric cancer proved by biopsies who received preoperative CEUS examination were involved in this study, and the CEUS result was compared with postoperative pathological findings. RESULTS: The normal gastric wall presented a one-layer structure in the portal venous phase and a three-layer structure in the arterial and equilibrium phase including a slightly hyper-enhanced inner layer, a hypo-enhanced intermediate layer, and a markedly hyper-enhanced outer layer, which corresponded histologically to the mucosal, submucosal, and muscular-serosal layer, respectively. The accuracy of transabdominal ultrasonography and CEUS in determining the T stage of gastric cancer was 72.9% and 88.1% respectively, and the difference was statistically significant (chi(2)=4.37, P=0.036). CONCLUSIONS: CEUS shows the normal gastric wall as a one- or a three-layer structure, which provides a theory base for CEUS in preoperative T-staging of gastric cancer. CEUS is a useful diagnostic method for preoperative T-staging of gastric cancer.

[Study on the induced differentiation of bone marrow-derived mesenchymal stem cells into inner ear hair cell-like cells in vitro].

OBJECTIVE: To investigate the feasibility of orientation-induced differentiation of rat bone marrow-derived mesenchymal stem cells (MSC) into cochlear hair cell-like cells in vitro. METHODS: MSC were extracted, cultured and identified. The organ of Corti was isolated from post natal day 1-day 3 Sprague-Dawley rats. Then MSC were co-cultured with organ of Corti for 14 days in vitro. The expression of hair cell markers (myosin VIIa, math1 and calretinin) was detected by immunocytochemical analyses and by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The differentiated cells were immunopositive for myosin VIIa, calretinin and math1. And RT-PCR results indicated that these differentiated cells expressed hair cell markers Math1 and myosin VIIa. CONCLUSIONS: MSC cultured in vitro could be successfully induced to differentiate into hair cell-like cells with hair cell molecular markers.

[Diagnostic value of double contrast-enhanced ultrasonography in preoperative staging of gastric cancer].

OBJECTIVE: To evaluate the clinical value of double contrast-enhanced ultrasonography using oral and intravenous contrast agents in preoperative staging of gastric cancer. METHODS: Sixty-two patients with biopsy-proven gastric cancer were enrolled into this study, and were examined by double contrast-enhanced gastric ultrasonography preoperatively. The results were compared with postoperative pathologic findings. RESULTS: The accuracy of oral contrast-enhanced gastric ultrasonography and double contrast-enhanced ultrasonography in determining the T stage of gastric cancer was 72.9% (T1: 66.7%, T2: 60.0%, T3: 76.9%, T4: 71.4%) and 88.1% (T1: 66.7%, T2: 80.0%, T3: 89.7%, T4: 100%), respectively, with a statistically significant difference between the two methods (P = 0.036). The sensitivity, specificity, accuracy and Youden index of oral contrast-enhanced gastric ultrasonography and double contrast-enhanced ultrasonography in assessment of lymph node metastasis were 74.5%, 66.7%, 72.9%, and 0.41 versus 89.4%, 75.0%, 86.4%, 0.76, respectively. No significant difference in the accuracy of assessment for lymph node metastasis was observed (P > 0.05). CONCLUSION: Double contrast-enhanced ultrasonography is useful for preoperative staging of gastric cancer, especially for T staging.

[Clinical effect of treatment with lipo-prostaglandin E1 on the patients with chronic glomerulonephritis].

OBJECTIVE: To investigate potential mechanism underlying lipo-prostaglandin E1 (PGE1) in treatment of patients with chronic glomerulonephritis. METHODS: The recommended dose of 20 microg lipo-PGE1 for treatment of chronic glomerulonephritis was as a daily dose by continuous intravenous infusion or intravenous injection for 4 weeks. The levels of interleukin-1 (IL- 1), tumor necrosis factor-alpha (TNF-alpha), blood urea nitrogen (BUN), serum creatinine (SCr) and clearance of creatinine (CCr) were measured before and after treatment. RESULTS: After treatment with lipo-PGE1, levels of IL-1, TNF-alpha, BUN and SCr were lower than those before treatment (all P<0.01), but CCr values were higher than those before treatment (P<0.05). CONCLUSION: Lipo-PGE1 can reduce the renal inflammatory response and improve the renal function in the patients with chronic glomerulonephritis.

[The application of ultrasonography to estimate blood vessel injury of upper limbs sustaining electric burns].

OBJECTIVE: To explore a new method in estimating extent and degree of arterial injury in upper limbs sustaining high tension electric burns. METHODS: Eighteen patients (twenty-four upper limbs) with high tension electricity injury were admitted from December 1998 to September 2002, The damaged limbs consisted of four parts: wrist wound part, 5 cm, 10 cm, 15 cm parts around wrist wound, where the radial and ulnar arteries were detected using B ultrasound and color WP Doppler examination. The changes of endangium, vessel diameter, thickness of the vessel wall and volume of blood flow were recorded respectively. The parameters of normal radial and ulnar arteries were also determined as normal control. RESULTS: B ultrasound and color WP Doppler examination showed that the endangium in radial and ulnar arteries become coarse, edema or exfoliation. The vessel wall was thicker than that of the normal control and the thickness was heterogeneity. The vessel wall could be necrosis in severe patient and the vessel cavity was stricture or beaded. Thrombosis or occlusion could occur at the site of severe injury area in vessel. The decrease in volume of blood flow was observed. The condition of the radial and ulnar arteries become well apart from 10 - 15 cm of wrist wound. CONCLUSIONS: The ultrasonography can be used to detect the changes in endangium, diameter, thickness of the vessel wall, blood flow volume in injury blood vessel caused by electric burn injury. It is helpful in judging the degree and extent of injury vessel and could be a safe, non-invasive diagnostic method and is worth popularizing.