Background: In addition to vascular endothelial cells, vascular smooth muscle cells (VSMCs) are subject to continuous shear stress because of blood circulation. The angiogenic properties of VSMCs in extracranial arteriovenous malformations (AVMs) may exceed those of normal blood vessels if the body responds more sensitively to mechanical stimuli. This study was performed to investigate the hypothesis that rapid angiogenesis may be achieved by mechanical shear stress. Methods: VSMCs were obtained from six patients who had AVMs and six normal controls. The target genes were set to angiopoietin-2 (AGP2), aquaporin-1 (AQP1), and transforming growth factor-beta receptor 1 (TGFBR1). Reverse-transcriptase polymerase chain reaction (RT-PCR) and real-time PCR were implemented to identify the expression levels for target genes. Immunofluorescence was also conducted. Results: Under the shear stress condition, mean relative quantity values of AGP2, AQP1, and TGFBR1 in AVM tissues were 1.927±0.528, 1.291±0.031, and 2.284±1.461 when compared with neutral conditions. The expression levels of all three genes in AVMs were higher than those in normal tissue except for AQP1 under shear stress conditions. Immunofluorescence also revealed increased staining of shear stress-induced genes in the normal tissue and in AVM tissue. Conclusions Shear stress made the VSMCs of AVMs more sensitive. Although the pathogenesis of AVMs remains unclear, our study showed that biomechanical stimulation imposed by shear stress may aggravate angiogenesis in AVMs.

No abstract available.
Humans ; Surgeons* ; Thyroid Gland* ; Thyroid Neoplasms*

PURPOSE: This study was performed to evaluate the long-term effects and safety of intratracheal (IT) transplantation of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) in neonatal hyperoxic lung injury at postnatal day (P)70 in a rat model. MATERIALS AND METHODS: Newborn Sprague Dawley rat pups were subjected to 14 days of hyperoxia (90% oxygen) within 10 hours after birth and allowed to recover at room air until sacrificed at P70. In the transplantation groups, hUCB-MSCs (5x10(5)) were administered intratracheally at P5. At P70, various organs including the heart, lung, liver, and spleen were histologically examined, and the harvested lungs were assessed for morphometric analyses of alveolarization. ED-1, von Willebrand factor, and human-specific nuclear mitotic apparatus protein (NuMA) staining in the lungs and the hematologic profile of blood were evaluated. RESULTS: Impaired alveolar and vascular growth, which evidenced by an increased mean linear intercept and decreased amount of von Willebrand factor, respectively, and the hyperoxia-induced inflammatory responses, as evidenced by inflammatory foci and ED-1 positive alveolar macrophages, were attenuated in the P70 rat lungs by IT transplantation of hUCB-MSCs. Although rare, donor cells with human specific NuMA staining were persistently present in the P70 rat lungs. There were no gross or microscopic abnormal findings in the heart, liver, or spleen, related to the MSCs transplantation. CONCLUSION: The protective and beneficial effects of IT transplantation of hUCB-MSCs in neonatal hyperoxic lung injuries were sustained for a prolonged recovery period without any long-term adverse effects up to P70.
Animals ; *Cord Blood Stem Cell Transplantation ; Ectodysplasins/metabolism ; Humans ; Hyperoxia/*pathology ; Lung/metabolism/pathology ; Lung Injury/pathology/*surgery ; *Mesenchymal Stem Cell Transplantation ; Models, Animal ; Nuclear Matrix-Associated Proteins/metabolism ; Rats ; Trachea/*transplantation ; von Willebrand Factor/metabolism

BACKGROUND: The underlying pathogenesis of fat embolism-induced acute lung injury (ALI) has not been elucidated. In the present study, the pathogenesis of fat embolism-induced ALI was probed in association with neutrophilic oxidative stress in oleic acid (OA)-induced ALI of S-D rats. METHODS: OA was injected intravenously to provoke ALI in experimental rats. Five hours later, indices of ALI were measured to confirm the role of the neutrophilic respiratory burst. The effect of an inhibition of phospholipase A2 (PLA2) was also evaluated. RESULTS: The accumulation of neutrophils in the lung due to OA caused increased neutrophilic oxidative stress in lung, which was ameliorated by mepacrine. What were the results from inhibition of PLA2. CONCLUSION: Excess neutrophilic oxidative stress contributes to OA-induced ALI, which is lessened by the inhibition of PLA2.
Acute Lung Injury ; Animals ; Embolism, Fat ; Lung ; Neutrophils ; Oleic Acid ; Oxidative Stress ; Phospholipases A2 ; Quinacrine ; Rats ; Respiratory Burst

PURPOSE: The prevalence rate of the BRAF mutation in papillary thyroid cancer (PTC) is as high as about 52 to 83% in Korea. Preoperative detection of BRAF mutation on fine needle aspiration cytology (FNAC) slides may help the surgeon make better therapeutic decisions. The present study aims to assess the feasibility of the mutant allele specific amplification (MASA) and restriction fragment length polymorphism (RFLP) method with using conventional FNAC slides and we also wanted to evaluate the clinical role of preoperatively detecting BRAF mutation. METHODS: We extracted the genomic DNA from 59 FNAC slides and performed direct sequencing (DS) for detecting BRAF mutation. We could use only 17 slides for the MASA method and 6 slides for the RFLP method due to the shortage of extracted DNA. Additionally, we retrospectively analyzed the cases for which a histological diagnosis could be made. RESULTS: Genomic DNA was extracted from 23 out of the 59 FNAC slides. The BRAF mutation status could be assessed via DS in 33 out of the 59 FNAC slides. The concordance between the MASA method and DS and the RFLP method and DS was 36.3% and 66.7% respectively. The positive and negative predictive value of the 13 indeterminate nodules was 87.5% and 20%, respectively. We could not find any association between the BRAF mutations and the alleged risk factors of PTC. CONCLUSION: We believe that the purity and the amount of the DNA template must be increased to detect BRAF mutation with using a FNAC slide. Preoperative detection of the BRAF mutation on a FNAC slide may refine the cytological diagnosis, but the application of assessing BRAF mutation as a prognostic marker is debatable.
Alleles ; Biopsy, Fine-Needle* ; Diagnosis* ; DNA ; Korea ; Methods ; Polymorphism, Restriction Fragment Length ; Prevalence ; Retrospective Studies ; Risk Factors ; Thyroid Gland* ; Thyroid Neoplasms*

We investigated the role of the central MAPK pathways in extra-territorial (referred) pain resulting from inflammation of the temporomandibular joint (TMJ). Experiments were carried out on male Sprague-Dawley rats weighing 220-280 g. Under anesthesia, these animals were injected with 50 microL of complete Freund's adjuvant (CFA) into the TMJ using a Hamilton syringe. In the control group, saline was injected into the TMJ. To identify the extent of inflammation of the TMJ, Evans blue dye (0.1%, 5 mg/kg) was injected intravenously at 1, 3, 6, 9, 12 and 15 days after CFA injection. The concentration of Evans blue dye in the extracted TMJ tissue was found to be significantly higher in the CFA-treated animals than in the saline-treated group. Air-puff thresholds in the vibrissa pad area were evaluated 3 days before and at 3, 6, 9, 12, 15 and 18 days after CFA injection into the TMJ. Referred mechanical allodynia was established at 3 days, remained until 12 days, and recovered to preoperative levels at 18 days after CFA injection. This referred mechanical allodynia was observed in contralateral side area. To investigate the role of central MAPK pathways, MAPK inhibitors (10 microg) were administrated intracisternally 9 days after CFA injection. SB203580, a p38 MAPK inhibitor, significantly attenuated referred mechanical allodynia, as compared with the vehicle group. PD98059, a MEK inhibitor, also reduced CFA-induced referred mechanical allodynia. These results suggest that TMJ inflammation produces extra-territorial mechanical allodynia, and that this is mediated by central MAPK pathways.
Anesthesia ; Animals ; Evans Blue ; Flavonoids ; Freund's Adjuvant ; Humans ; Hyperalgesia ; Imidazoles ; Inflammation ; Male ; p38 Mitogen-Activated Protein Kinases ; Pain, Referred ; Pyridines ; Rats ; Rats, Sprague-Dawley ; Syringes ; Temporomandibular Joint

The present study investigated the role of ERK in the onset of mechanical and cold allodynia in a rat model of compression of the trigeminal ganglion by examining changes in the air-puff thresholds and number of scratches following the intracisternal injection of PD98059, a MEK inhibitor. Male Sprague Dawley rats weighing between 250 and 260 g were used. Under anesthesia, the rats were mounted onto a stereotaxic frame and received 4% agar (10 microl) solution to compress the trigeminal ganglion. In the control group, the animals were given a sham operation without the application of agar. Changes in behavior were examined at 3 days before and at 3, 7, 10, 14, 17, 21, 24, 30, and 40 days after surgery. Compression of the trigeminal ganglion significantly decreased the air-puff thresholds. Mechanical allodynia was established within 3 days and persisted over postoperative day 24. To evaluate cold allodynia, nociceptive scratching behavior was monitored after acetone application on the vibrissa pad of the rats. Compression of the trigeminal ganglion was found to produce significant cold allodynia, which persisted for more than 40 days after surgery. On postoperative day 14, the intracisternal administration of 1 microg or 10 microg of PD98059 in the rat model significantly decreased the air-puff thresholds on both the ipsilateral and contralateral side. The intracisternal administration of 10 microg of PD98059 also significantly alleviated the cold allodynia, compared with the vehicle-treated group. These results suggest that central ERK plays an important role in the development of mechanical and cold allodynia in rats with compression of the trigeminal ganglion and that a targeted blockade of this pathway is a potential future treatment strategy for trigeminal neuralgia-like nociception.
Acetone ; Agar ; Anesthesia ; Animals ; Cold Temperature ; Flavonoids ; Humans ; Hyperalgesia ; Male ; Nociception ; Rats ; Rats, Sprague-Dawley ; Salicylamides ; Trigeminal Ganglion ; Trigeminal Neuralgia

Intestinal ischemia/reperfusion (I/R) is one of common causes of acute lung injury (ALI). Early and accurate diagnosis of patients who are like to develop serious acute respiratory distress syndrome (ARDS) would give a therapeutic advantage. Ferritin and heme oxygenase-1 (HO-1) are increased by oxidative stress and are potential candidates as a predictive biomarker of ARDS. However, the mechanisms responsible for the increases of ferritin and HO-1, and their relationship to ALI, are unclear. In order to elucidate the interactions between ferritin and HO-1, we studied the changes in ferritin and HO-1 levels in serum and bronchoalveolar lavage (BAL) fluid after intestinal I/R injury in rats. Leukocyte number and protein contents in BAL fluid were elevated following I/R, and the increases were attenuated by mepacrine pretreatment. Both serum ferritin and HO-1 concentrations were progressively elevated throughout the 3 h observation period. Mepacrine pretreatment attenuated the increase of serum and BAL fluid ferritin concentrations, but did not suppress the increase of serum HO-1. Moreover, BAL fluid HO-1 levels did not change after I/R or after mepacrine pretreated I/R compared with sham rats. Unlike ferritin, HO-1 levels are not exactly matched with the ALI. Therefore, there might be a different mechanism between the changes of ferritin and HO-1 in intestinal I/R-induced ALI model.
Acute Lung Injury ; Animals ; Bronchoalveolar Lavage ; Ferritins ; Heme ; Heme Oxygenase-1 ; Humans ; Imidazoles ; Leukocyte Count ; Lung ; Lung Injury ; Nitro Compounds ; Oxidative Stress ; Quinacrine ; Rats ; Respiratory Distress Syndrome, Adult ; Salicylamides

BACKGROUND: For unknown reasons, the serum ferritin concentrations are higher in patients with acute lung injury. A pretreatment with aspirin reduces the acute lung injury in rats subjected severe hemorrhage, and increases the rate of ferritin synthesis in vitro. This study investigated the effect of aspirin on the serum ferritin changes in rats subjected to severe hemorrhage. METHODS: Hemorrhagic shock was induced by withdrawing blood (20 ml/kg of B.W.) through the femoral artery for 5 min. The rats were pretreated with aspirin (10 mg/kg, i.v.) 30 min before hemorrhage. RESULTS: The protein content and leukocyte count in the bronchoalveolar lavage fluid, lung tissue myeloperoxidase activities were significantly higher after hemorrhage. The aspirin pretreatment prevented these changes. The serum and lavage fluid ferritin concentrations were elevated higher after hemorrhage. These were also attenuated by the aspirin pretreatment. CONCLUSION: The changes in the serum and lung lavage ferritin level might be closely related to the severity of hemorrhage?induced acute lung injury. Therefore, the serum and lavage ferritin concentrations can be a useful biomarker for patients with precipitating conditions.
Acute Lung Injury* ; Animals ; Aspirin* ; Bronchoalveolar Lavage ; Bronchoalveolar Lavage Fluid ; Femoral Artery ; Ferritins ; Hemorrhage* ; Humans ; Leukocyte Count ; Lung ; Peroxidase ; Rats* ; Shock, Hemorrhagic ; Therapeutic Irrigation

BACKGROUND: For unknown reasons, the serum ferritin concentrations are higher in patients with acute lung injury. A pretreatment with aspirin reduces the acute lung injury in rats subjected severe hemorrhage, and increases the rate of ferritin synthesis in vitro. This study investigated the effect of aspirin on the serum ferritin changes in rats subjected to severe hemorrhage. METHODS: Hemorrhagic shock was induced by withdrawing blood (20 ml/kg of B.W.) through the femoral artery for 5 min. The rats were pretreated with aspirin (10 mg/kg, i.v.) 30 min before hemorrhage. RESULTS: The protein content and leukocyte count in the bronchoalveolar lavage fluid, lung tissue myeloperoxidase activities were significantly higher after hemorrhage. The aspirin pretreatment prevented these changes. The serum and lavage fluid ferritin concentrations were elevated higher after hemorrhage. These were also attenuated by the aspirin pretreatment. CONCLUSION: The changes in the serum and lung lavage ferritin level might be closely related to the severity of hemorrhage?induced acute lung injury. Therefore, the serum and lavage ferritin concentrations can be a useful biomarker for patients with precipitating conditions.
Acute Lung Injury* ; Animals ; Aspirin* ; Bronchoalveolar Lavage ; Bronchoalveolar Lavage Fluid ; Femoral Artery ; Ferritins ; Hemorrhage* ; Humans ; Leukocyte Count ; Lung ; Peroxidase ; Rats* ; Shock, Hemorrhagic ; Therapeutic Irrigation