BACKGROUND: The nucleus tractus solitarius (NTS), the region of the brain stem in which primary baroreceptor afferents teminate, is critically important in the normal regulation of arterial pressure (AP). In the NTS, excitatory amino acids such as L-glutamate serve as the main neurotransmitter in the regulation of AP. However, the function of GABA in the NTS has not been established. To test the function of GABA, we applied GABAergic agents to the NTS. METHODS: The experiments were conducted on adult male Sprague-Dawley rats weighing 300-500g. A cannula (PE-50 tubing filled with heparinized saline) was inserted into the femoral artery for recording of AP and heart rate(HR). Another cannula was inserted into the femoral vein for administration of nitroprusside or phenylephrine. After rats were placed on a sterotaxic instrument, the dorsal surface of the medulla was exposed, and with the aid of a surgical microscope, the NTS was visualized. Drug injections were made into the NTS using single- or three-barreled grass micropipettes pulled to an outer diameter of 80-100(micro)m and connected to a 1(micro)l Hamilton syringe. RESULTS: The follwing results were obtained in this experiment. Injection into the NTS of 10 or 20 nmol nipecitic acid, a selective inhibitor of GABA untake, produced an increase in AP. The pressor responses evoked by two doses of nipecotic acid were not significantly different. Injection of GABA(A) agonist, musciml(5 pmol in 80 nl artificial CSF) and GABA(B) agonist, baclofen (20 pmol in 80 nl) into the NTS of urethane-anesthetized rats prodused an increase in AP of 16.6+/-1.3 and 27.6+/-1.5 mmHg, respectively. Thus the pressor response to GABA(B) agonist was greater than to GABA(A) agonist. On the other hand, microinjection of GABA(A) antagonist, bicuculline and GABA(B) antagonist, phaclofen into the NTS decreased AP by approximately 13.4+/-1.0 and 20.9+/- mmHg, respectively. Thus injection of nipecotic acid into the NTS was greater in control group compared with the muscimiol or baclofen groups. The AP changes caused by i.v. injection of nitroprusside or phenylephrine were smallest in control group and greatest in the baclofen group. When calculated as baroreflex sensitivity, the change was greatest in control group and smallest in the baclofen group. CONCLUSION: From these results it was concluded that GABA in the NTS plays an important role in the regulation of AP, especially through GABA(B) receptors, and have an inhibitory effect on baroreceptor reflex.
Adult ; Animals ; Arterial Pressure ; Baclofen ; Baroreflex ; Bicuculline ; Blood Pressure ; Brain Stem ; Catheters ; Excitatory Amino Acids ; Femoral Artery ; Femoral Vein ; GABA Agents ; gamma-Aminobutyric Acid* ; Glutamic Acid ; Hand ; Heart ; Heparin ; Humans ; Male ; Microinjections ; Neurotransmitter Agents ; Nitroprusside ; Phenylephrine ; Poaceae ; Pressoreceptors ; Rats ; Rats, Sprague-Dawley ; Solitary Nucleus* ; Syringes

BACKGROUND/AIMS: Transarterial chemoembolization (TACE) has been reported to be one of the useful palliative treatments in patients with unresectable hepatocelluar carcinoma. However, Bile duct injuries following TACE have been reported occasionally. In this study, we intended to clarify the incidence, pathogenic mechanisms and clinical implications of bile duct injuries following TACE. METHODS: A total of 950 consecutive patients with hepatocellular carcinoma (HCC) were subjected. 807 patients were treated with TACE. The remaining 143 were treated with transarterial chemoinfusion (TACI) of cisplatin. RESULTS: None of 143 HCC patients treated with TACI revealed to have any ischemic biliary injury radiologically. In contrast, out of 807 with TACE, 17 (2%) appeared to have biliary complications. Twelve out of 17 (71%) had bilomas at subcapsular area, three out of 17 (18%) had focal strictures at common hepatic duct or common bile duct with marked dilatation of intrahepatic bile ducts and two out of 17 (11%) had diffuse mild dilatation of intrahepatic bile ducts. Interestingly, two (17%) out of 12 bilomas were found at the lobe which was not embolized with Gelfoam. The median sessions of TACE to the occurrences of focal strictures tended to be longer compared with those of bilomas (median: 6 vs. 2.5; p=0.08). All three patients with focal strictures and four (33%) out of 12 patients with bilomas were associated with serious bacterial infections at presentation. CONCLUSIONS: Biloma seems to be caused by lipiodol rather than Gelfoam; focal strictures of large bile ducts by Gelfoam. It is suggested that adjustments of the amounts of lipiodol or Gelfoam and the sites or embolization may be required to reduce the ischemic biliary injuries following TACE.
Bacterial Infections ; Bile Ducts* ; Bile Ducts, Intrahepatic ; Bile* ; Carcinoma, Hepatocellular ; Cisplatin ; Common Bile Duct ; Constriction, Pathologic ; Dilatation ; Ethiodized Oil ; Gelatin Sponge, Absorbable ; Hepatic Duct, Common ; Humans ; Incidence ; Palliative Care

BACKGROUND: Most lung cancer patients receive systemic chemotherapy at an advanced stage disease. Cisplatin-based chemotherapy is the main regimen for treating advanced lung cancer. Recently, autophagy has become an important mechanism of cellular adaptation under starvation or cell oxidative stress. The purpose of this study was to determine whether or not autophagy can occurred in cisplatin-treated lung cancer cells. METHODS: H460 cells were incubated with RPMI 1640 and treated in 5 micrometer or 20 micrometer cisplatin concentrations at specific time intervals. Cells surviving cisplatin treatment were measured and compared using an MTT cell viability assay to cells that underwent apoptosis with autophagy by nuclear staining, apoptotic or autophagic related proteins, and autophagic vacuoles. The development of acidic vascular organelles was using acridine orange staining and fluorescent expression of GFP-LC3 protein in its transfected cells was observed to evaluate autophagy. RESULTS: Lung cancer cells treated with 5 micrometer cisplatin-treated were less sensitive to cell death than 20 micrometer cisplatin-treated cells in a time-dependent manner. Nuclear fragmentation at 5 micrometer was not detected, even though it was discovered at 20 micrometer. Poly (ADP-ribose) polymerase cleavages were not detected in 5 micrometer within 24 hours. Massive vacuolization in the cytoplasm of 5 micrometer treated cells were observed. Acridine orange stain-positive cells was increased according in time-dependence manner. The autophagosome-incorporated LC3 II protein expression was increased in 5 micrometer treated cells, but was not detected in 20 micrometer treated cells. The expression of GFP-LC3 were increased in 5 micrometer treated cells in a time-dependent manner. CONCLUSION: The induction of autophagy occurred in 5 micrometer dose of cisplatin-treated lung cancer cells.
Acridine Orange ; Apoptosis ; Autophagy ; Cell Death ; Cell Survival ; Cisplatin ; Cytoplasm ; Humans ; Lung ; Lung Neoplasms ; Organelles ; Oxidative Stress ; Proteins ; Starvation ; Vacuoles

BACKGROUND: Most lung cancer patients receive systemic chemotherapy at an advanced stage disease. Cisplatin-based chemotherapy is the main regimen for treating advanced lung cancer. Recently, autophagy has become an important mechanism of cellular adaptation under starvation or cell oxidative stress. The purpose of this study was to determine whether or not autophagy can occurred in cisplatin-treated lung cancer cells. METHODS: H460 cells were incubated with RPMI 1640 and treated in 5 micrometer or 20 micrometer cisplatin concentrations at specific time intervals. Cells surviving cisplatin treatment were measured and compared using an MTT cell viability assay to cells that underwent apoptosis with autophagy by nuclear staining, apoptotic or autophagic related proteins, and autophagic vacuoles. The development of acidic vascular organelles was using acridine orange staining and fluorescent expression of GFP-LC3 protein in its transfected cells was observed to evaluate autophagy. RESULTS: Lung cancer cells treated with 5 micrometer cisplatin-treated were less sensitive to cell death than 20 micrometer cisplatin-treated cells in a time-dependent manner. Nuclear fragmentation at 5 micrometer was not detected, even though it was discovered at 20 micrometer. Poly (ADP-ribose) polymerase cleavages were not detected in 5 micrometer within 24 hours. Massive vacuolization in the cytoplasm of 5 micrometer treated cells were observed. Acridine orange stain-positive cells was increased according in time-dependence manner. The autophagosome-incorporated LC3 II protein expression was increased in 5 micrometer treated cells, but was not detected in 20 micrometer treated cells. The expression of GFP-LC3 were increased in 5 micrometer treated cells in a time-dependent manner. CONCLUSION: The induction of autophagy occurred in 5 micrometer dose of cisplatin-treated lung cancer cells.
Acridine Orange ; Apoptosis ; Autophagy ; Cell Death ; Cell Survival ; Cisplatin ; Cytoplasm ; Humans ; Lung ; Lung Neoplasms ; Organelles ; Oxidative Stress ; Proteins ; Starvation ; Vacuoles

BACKGROUND: Biomarkers for cancer have several potential clinical uses, including the following: early cancer detection, monitoring for recurrence prognostication, and risk stratification. However, no biomarker has been shown to have adequate sensitivity and specificity. Many investigators have tried to validate biomarkers for the early detection and recurrence of lung cancer. To evaluate plasma G-CSF as such a biomarker, protein levels were measured and were found to correlate with the clinicopathological features of primary lung tumors. METHODS: Between December 2006 and May 2008, 100 patients with histologically-validated primary lung cancer were enrolled into this study. To serve as controls, 127 healthy volunteers were enrolled into this study. Plasma G-CSF levels were measured in lung cancer patients using the sandwich ELISA system (R & D inc.) prior to treatment. RESULTS: The mean plasma G-CSF levels were 12.2+/-0.3 pg/mL and 46.0+/-3.8 pg/mL (mean+/-SE) in the normal and in the cancer groups, respectively. In addition, plasma G-CSF levels were higher in patients with early lung cancer than in healthy volunteers (p<.001). Plasma G-CSF levels were higher in patients who were under 65 years old or smokers. Within the cancer group, plasma G-CSF levels were higher in patients with non small cell lung cancer than in patients with small cell lung cancer (p<.05). Overall, plasma G-CSF levels were shown to increase dependent upon the type of lung cancer diagnsosed. In the order from highest to lowest, the levels of plasma G-CSF tended to decrease in the following order: large cell carcinoma, squamous cell carcinoma, adenocarcinoma, and bronchioloalveolar carcinoma. Plasma G-CSF levels tended to be higher in patients with advanced TNM stage than in localized TNM stage (I, II Adenocarcinoma ; Adenocarcinoma, Bronchiolo-Alveolar ; Adrenal Glands ; Biomarkers ; Carcinoma, Large Cell ; Carcinoma, Squamous Cell ; Enzyme-Linked Immunosorbent Assay ; Granulocyte Colony-Stimulating Factor ; Humans ; Lung ; Lung Neoplasms ; Neoplasm Metastasis ; Plasma ; Recurrence ; Research Personnel ; Small Cell Lung Carcinoma

BACKGROUND: Autophagy is an important adaptive mechanism in normal development and in response to changing environmental stimuli in cancer. Previous papers have reported that different types of cancer underwent autophagy to obtain amino acids as energy source of dying cells in nutrient-deprived conditions. However, whether or not autophagy in the process of lung cancer causes death or survival is controversial. Therefore in this study, we investigated whether nutrient deprivation induces autophagy in human H460 lung cancer cells. METHODS: H460, lung cancer cells were incubated in RPMI 1640 medium, and the starved media, which are BME and RPMI media without serum, including 2-deoxyl-D-glucose according to time dependence. To evaluate the viability and find out the mechanism of cell death under nutrient-deprived conditions, the MTT assay and flow cytometry were done and analyzed the apoptotic and autophagic related proteins. It is also measured the development of acidic vascular organelles by acridine orange. RESULTS: The nutrient-deprived cancer cell is relatively sensitive to cell death rather than normal nutrition. Massive cytoplasmic vacuolization was seen under nutrient-deprived conditions. Autophagic vacuoles were visible at approximately 12 h and as time ran out, vacuoles became larger and denser with the increasing number of vacuoles. In addition, the proportion of acridine orange stain-positive cells increased according to time dependence. Localization of GFP-LC3 in cytoplasm and expression of LC-3II and Beclin 1 were increased according to time dependence on nutrient-deprived cells. CONCLUSION: Nutrient deprivation induces cell death through autophagy in H460 lung cancer cells.
Acridine Orange ; Amino Acids ; Autophagy ; Cell Death ; Cytoplasm ; Flow Cytometry ; Humans ; Lung Neoplasms ; Malnutrition ; Organelles ; Proteins ; Vacuoles