Recently, ultrasound-targeting microbubble destruction has been employed in molecular gene therapy, and a new potent nonviral gene transfer method known as 'sonoporation' has been developed. We investigated the efficiency of sonoporation toward growth inhibition of human gingival squamous carcinoma cell line, Ca9-22, in vitro and in vivo. The cytotoxicity of bleomycin (BLM) was investigated using flow-cytometric analysis and Hoechst's staining in vitro assay systems. We found that the delivery of BLM by sonoporation induced cytotoxic effect toward Ca9-22 cells in vitro. Our in vivo results showed that tumors nearly disappeared in Ca9-22 cell-implanted nude KSN/slc mice treated with a low dose of BLM followed by sonoporation during the 4-week experimental period. Histological analysis revealed that the cytotoxic effect was mainly apoptosis. We previously reported that the cytolethal distending toxin B (cdtB) from Actinobacillus actinomycetemcomitans, a periodontopathic bacterium, is responsible for cell cycle arrest and apoptosis in vitro. Thus, we used sonoporation to transfect a cdtB-expressing plasmid into Ca9-22 cells and examined cell viability in vitro and in vivo. We found that an administration of cdtB-expressing plasmid followed by sonoporationinduced marked growth inhibition of Ca9-22 cells and apoptotic cells were also observed in vitro and in vivo. These findings suggest that local administration of cytotoxic agents with sonoporation is a useful method for molecular cancer therapy.

The parotid glands of 228 Japanese human cadavers were examined to determine the incidence and histological features of accessory parotid glands. The incidence was found to be 56% with no differences between right and left sides or between sexes. Thirty parotid glands and their associated accessory glands were examined histologically: eight of these accessory glands were found to be mixed secretory glands (i.e., containing both serous and mucous acini). Thus, the pattern of differentiation of a significant fraction of accessory glands differs from that of the main parotid gland: it appears that mixed acini, present in the early stages of development, persist into later life, and their presence may be related to tumors developing at these sites.

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The MW of HA in SF from patients with TMD is decreased.