Praeruptorin-B Inhibits 12-O-Tetradecanoylphorbol-13-Acetate-Induced Cell Invasion by Targeting AKT/NF-κB via Matrix Metalloproteinase-2/-9 Expression in Human Cervical Cancer Cells.

BACKGROUND/AIMS: Praeruptorins, a seselin-type coumarin, possess anti-inflammatory and antitumor promoting properties. However, molecular mechanisms through which Praeruptorin-B (Pra-B) exerts an antimetastatic effect on cervical cancer cells remain unclear. METHODS: Cell viability was examined using the MTT assay, whereas cell migration and invasion were examined using the Boyden chamber assay. Western blotting and RT-PCR were performed to investigate the inhibitory effect of Pra-B on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced matrix metalloproteinase-2/-9 (MMP-2/-9) expression in HeLa cells. The findings of the luciferase assay confirmed the inhibitory effect of Pra-B on TPA-induced transcriptional activity of MMP2/-9 in HeLa cells. RESULTS: Pra-B inhibited TPA-induced metastatic ability of human cervical cancer cells without any significant toxicity. Pra-B suppressed TPA-induced mRNA and protein expression and transcriptional activity of MMP-2/-9 in HeLa cells. Furthermore, Pra-B inhibited AKT phosphorylation but did not affect the MAPK pathway. Cotreatment of HeLa cells with TPA plus Pra-B or LY294002 (a PI3K inhibitor) reduced cell invasion and MMP-2/-9 expression and transcriptional activity. In addition, Pra-B attenuated TPA-induced nuclear translocation of NF-κB-p65/-p50, which reduced Ikk-α phosphorylation in HeLa cells. Cotreatment of HeLa cells with TPA plus Pra-B or LY294002 reduced NF-κB nuclear translocation. CONCLUSION: These results suggested that Pra-B-mediated inhibition of TPA-induced cell metastasis involved the suppression of p-AKT/NF-κB via MMP-2/-9 expression in HeLa cells. Pra-B can be a potential antimetastatic agent against cervical cancer.

Synergistic effect of fisetin combined with sorafenib in human cervical cancer HeLa cells through activation of death receptor-5 mediated caspase-8/caspase-3 and the mitochondria-dependent apoptotic pathway.

Combining antitumor agents with bioactive compounds is a potential strategy for improving the effect of chemotherapy on cancer cells. The goal of this study was to elucidate the antitumor effect of the flavonoid, fisetin, combined with the multikinase inhibitor, sorafenib, against human cervical cancer cells in vitro and in vivo. The combination of fisetin and sorafenib synergistically induced apoptosis in HeLa cells, which is accompanied by a marked increase in loss of mitochondrial membrane potential. Apoptosis induction was achieved by caspase-3 and caspase-8 activation which increased the ratio of Bax/Bcl-2 and caused the subsequent cleavage of PARP level while disrupting the mitochondrial membrane potential in HeLa cells. Decreased Bax/Bcl-2 ratio level and mitochondrial membrane potential were also observed in siDR5-treated HeLa cells. In addition, in vivo studies revealed that the combined fisetin and sorafenib treatment was clearly superior to sorafenib treatment alone using a HeLa xenograft model. Our study showed that the combination of fisetin and sorafenib exerted better synergistic effects in vitro and in vivo than either agent used alone against human cervical cancer, and this synergism was based on apoptotic potential through a mitochondrial- and DR5-dependent caspase-8/caspase-3 signaling pathway. This combined fisetin and sorafenib treatment represents a novel therapeutic strategy for further clinical developments in advanced cervical cancer.

Structural and electrical characterization of ohmic contacts to graphitized silicon carbide.

Titanium was deposited onto silicon carbide (6H-SiC) using the 248 nm line of an excimer laser in a vacuum of 10(-6) Torr, and ohmic contacts were formed by annealing the structure at approximately 1000 degrees C. Further anneals between 1350 and 1430 degrees C did not degrade the formed contacts, and Raman analysis confirmed that sublimation of silicon from the near surface layers of the silicon carbide between the contact pads resulted in graphene formation after 5 min, 1428 degrees C anneals. The graphene formation was accompanied by a significant enhancement of ohmic behavior, and, it was found to be sensitive to the temperature ramp-up rate and annealing time. High-resolution transmission electron microscopy showed that the interface between the metal and silicon carbide remained sharp and free of macroscopic defects even after 30 min, 1430 degrees C anneals. The interface was determined to be carbon rich by elemental analysis, which indicates metal carbide formation. The potential of this approach for achieving ohmic contacts and graphene formation on silicon carbide substrates is discussed. A mechanism for the sequential formation of ohmic contacts then graphene is proposed.

A new method using epidural catheters in the reconstruction of lacrimal drainage.

Lacrimal outflow obstruction after severance of the duct is a common problem in facial trauma. Conventional treatments include external dacryocystorhinostomy, endoscopic-assisted dacryocystorhinostomy, conjunctivorhinostomy, and a Jones tube bypass. However, the disadvantages of these methods are that the procedures are complicated and there is a high rate of recurrence. From April 2000 to March 2003, seven patients with epiphora after facial trauma had their lacrimal ducts drained with epidural catheters. The V-M shape incision was used with an epidural catheter placed as a stent for six months. After removal of the tube, all patients recovered fully from the epiphora during the follow-up period, and there were no complications.

Establishment of an immortalized porcine granulosa cell line (PGV) and the study on the potential mechanisms of PGV cell proliferation.

In order to establish an immortalized granulosa cell line and to investigate the potential mechanisms of immortalized cell proliferation, simian virus (SV) 40 was used to infect porcine granulosa cells from small follicles (1-2 mm in diameter), and one colony was selected after four weeks of culture. The colony was digested with trypsin and the cells were cultured for more than 300 days (named PGV). The SV40 large T antigen gene and its products were confirmed in immortalized cells by Southern blotting and immunohistochemistry. Progesterone production was not detected in the conditioned culture media with follicle-stimulating hormone (FSH) and forskolin, possibly due to the lack of P450scc gene transcription as examined by Northern blotting. PGV cells responded significantly to the stimulation of sera (fetal bovine and horse sera) and protein kinase C (PKC) stimulators (PMA and OAG), while PKC inhibitors (staurosporine and calphostin C) blocked both sera and PKC stimulation. Phospholipase C (PLC) and phosphatidic acid phosphatase (PAP) inhibitors (U73122 and propranolol) significantly reduced PGV cell proliferation, while PMA restored PLC and PAP inhibition. These data suggest that diacylglycerol (DAG) is produced in PGV cells by PLD as well as by PLC, and that DAG then activates PKC stimulating the PGV cell cycle through yet unknown mechanisms. Thus, an immortalized granulosa cell line is very useful to study granulosa cells in vitro, as the cells are homogeneous and are a functionally defined population.

Tensor fasciae latae combined with tangentially split vastus lateralis musculocutaneous flap for the reconstruction of pressure sores.

Pressure sores in the ischial and the trochanteric regions are usually encountered in long-term bedridden and wheelchair-dependent patients. A number of techniques have been developed for the reconstruction of pressure sores. Tensor fasciae latae musculocutaneous flap has been extensively employed to close the trochanteric defect. Despite its utility of having a constant pedicle and proximal bulky muscle, the relative shortness of the flap and insufficient padding in the distal portion limit the applications for distant locations of pressure sores. From January 2001 to December 2003, 8 patients with ischial and trochanteric pressure sores underwent tensor fasciae latae reconstruction in combination with tangentially split vastus lateralis muscle. The descending branches of the lateral circumflex femoral arteries were also included in these flaps. All of the procedures have been successful, and no flap necrosis has been observed. Sufficient bulk of the flap and reliable distal skin paddle constitute the advantages of this flap.

Intercellular calcium waves mediate preferential cell growth toward the wound edge in polarized hepatic cells.

During liver regeneration, hepatocytes sense the damage and initiate proliferation of the quiescent cells through poorly understood mechanisms. Here, we have used cultured hepatic cells to study the roles played by intercellular calcium in mediating wound-healing processes. Well-differentiated and polarized Hep-G2 cells repaired an experimentally induced wound by induction of cell divisions. The resulting cellular growth did not occur evenly across the healing cell lawn; instead, proliferations were three times more active within 150-200 microm from the wound edge than further away; this periwound preferential cell growth was not observed in the poorly differentiated and/or nonpolarized cells. We have provided experimental evidence demonstrating that the wounding procedure itself could elicit a propagating calcium wave, and interestingly, blocking this injury-associated intercellular calcium communication could effectively inhibit the biased cell growth along the margin of the wound. A photolithography-based patterned cell culture system was employed to help delineate the mechanisms underlying this type of calcium signaling. In conclusion, our results suggested that intercellular communications via propagating calcium waves coordinate regenerative cell proliferations in response to hepatic tissue losses.

Holoprosencephaly and cyclopia visualized by two- and three-dimensional prenatal ultrasound.

We report on the ultrasound (US) detection of holoprosencephaly with cyclopia at a gestational age of 16 weeks. The sonographic diagnosis was based on the intracranial finding of fused thalami with no visible midline structures and facial abnormalities, including cyclopia and proboscis. We evaluated the fetal face by 3-dimensional (3D) transabdominal US and were able to identify the cyclopia below the proboscis. These findings are characteristic of alobar holoprosencephaly. With the informed consent of the patient, the pregnancy was terminated by prostaglandin induction after proper counseling. Chromosome study of the abortus revealed a normal karyotype (46, XY). Postmortem examination of the abortus confirmed the presence of cyclopia and a proboscis. The use of 3D prenatal US made additional diagnostic images possible.