Ginseng's major active components, ginsenosides, have been known to show anti-cancer, neuroprotective, and anti-inflammatory activities. Ultrasonication processed Panax ginseng berry extract (UGB) contains various ginsenosides. The components are different from Panax ginseng berry extract (GBE). This study was aimed to investigate the cytotoxic mechanism of UGB in HepG2 cells, human hepatocellular carcinoma cell line. HepG2 cells were treated with UGB (0, 10, 20 μg/ml). Cell growth and cellular apoptosis were evaluated by MTT assay and Annexin V/Pi staining, respectively. Intracellular Reactive oxygen species (ROS) levels were also determined by 2', 7'-dichlorofluorescin diacetate (DCFDA) staining. The expressions of Bax, Bcl-2 and caspase-3, the apoptotic markers, were evaluated by Western Blot. UGB dose-dependently inhibited cell growth and induced apoptotic cell death. Intracellular ROS levels were increased. UGB increased the expression of the cleaved form of caspase-3. Furthermore, UGB induced apoptosis of HepG2 cells through Bax activation and Bcl-2 inhibition. In conclusion, UGB induced apoptosis through an intrinsic pathway in HepG2 cells suggesting that UGB might play a role as a novel substance for anti-cancer effect.
Red blood cell (RBC) deformability is greatly affected by the osmolality, and maximum deformability, which is determined as maximal elongation index (EI max ), is usually observed in isotonic conditions at high shear stresses (>20 Pa). Therefore, we examined osmotic RBC deformability over a range of shear stresses (0.5-20 Pa). We found that the RBC deformability at low shear stresses (1-3 Pa) was maximum in hypotonic conditions (225-250 mOsm/kg H 2 O), which is slightly lower than the normal range of osmolality in plasma (290-310 mOsm/kg H 2 O). The phenomenon that O max (the osmolality at EI max ) is dependent on applied shear stress could play an important role in microcirculation in which osmolality varies widely.
A 2.7 kg, 11-year-old, castrated male Maltese dog was presented for evaluation of a 2-year history of intractable coughing, dyspnoea and cyanosis. A diagnosis of tracheal collapse with myxomatous mitral valve disease was made on the basis of inspiratory and expiratory thoracic radiographs, fluoroscopy and echocardiography. Measurement for stent size selection was performed on thoracic radiographs. A 10 mm (diameter) × 70 mm (length) self-expanding double-wire woven uncovered nitinol stent was used for intratracheal implantation and was deployed under fluoroscopic guidance. On thoracic radiography seven days after surgery, the position of the stent remained unchanged. On presentation six months after surgery, the owner reported that the dog was doing well without medical management. Although studies of various intraluminal stents have been reported in dogs, to the authors' knowledge, use of a double-wire woven uncovered nitinol stent has not been reported previously for the management of a dog with tracheal collapse. Since this particular type of stent with unfixed individual cells provided proper airway patency without stent fracture in the dog in this report, this stent might be used as an alternative to other commercially available nitinol stents in cases of thoracic inlet collapse.
P2 receptors are membrane-bound receptors for extracellular nucleotides such as ATP and UTP. P2 receptors have been classified as ligand-gated ion channels or P2X receptors and G protein-coupled P2Y receptors. Recently, purinergic signaling has begun to attract attention as a potential therapeutic target for a variety of diseases especially associated with gastroenterology. This study determined the ATP and UTP-induced receptor signaling mechanism in feline esophageal contraction. Contraction of dispersed feline esophageal smooth muscle cells was measured by scanning micrometry. Phosphorylation of MLC20 was determined by western blot analysis. ATP and UTP elicited maximum esophageal contraction at 30 s and 10 μM concentration. Contraction of dispersed cells treated with 10 μM ATP was inhibited by nifedipine. However, contraction induced by 0.1 μM ATP, 0.1 μM UTP and 10 μM UTP was decreased by U73122, chelerythrine, ML-9, PTX and GDPβS. Contraction induced by 0.1 μM ATP and UTP was inhibited by Gαi3 or Gαq antibodies and by PLCβ1 or PLCβ3 antibodies. Phosphorylated MLC20 was increased by ATP and UTP treatment. In conclusion, esophageal contraction induced by ATP and UTP was preferentially mediated by P2Y receptors coupled to Gαi3 and G q proteins, which activate PLCβ1 and PLCβ3. Subsequently, increased intracellular Ca2+ and activated PKC triggered stimulation of MLC kinase and inhibition of MLC phosphatase. Finally, increased pMLC20 generated esophageal contraction.
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