The Effect of Epigallocatechin-3-gallate on HIF-1alpha and VEGF in Human Lung Cancer Cell Line / 결핵및호흡기질환

BACKGROUND: Epigallocatechin-3-gallate (EGCG) is the major catechin in green tea, and has shown antiproliferative, antiangiogenic, antimetastatic and cell cycle pertubation activity in various tumor models. Hypoxia can be induced because angiogenesis is insufficient for highly proliferating cancer. Hypoxia-inducible factor-1alpha (HIF-1alpha) and its downstream target, vascular endothelial growth factor (VEGF), are important for angiogenesis, tumor growth and metastasis. The aim of this study was to determine how hypoxia could cause changes in the cellular phenomena and microenvironment in a non-small cell culture system and to examine the effects of EGCG on a HIF-1alpha and VEGF in A549 cell line. METHODS: A549 cells, a non-small cell lung cancer cell line, were cultured with DMEM and 10% fetal bovine serum. A decrease in oxygen tension was induced using a hypoxia microchamber and a CO2-N2 gas mixture. Gas analysis and a MTT assay were performed. The A549 cells were treated with EGCG (0, 12.5, 25, 50 micromol/L), and then examined by real-time-PCR analysis of HIF-1alpha, VEGF, and beta-actin mRNA. RESULTS: Hypoxia reduced the proliferation of A549 cells from normoxic conditions. EGCG inhibited HIF-1alpha transcription in A549 cells in a dose-dependent manner. Compared to HIF-1alpha, VEGF was not inhibited by EGCG. CONCLUSION: HIF-1alpha can be inhibited by EGCG. This suggests that targeting HIF-1alpha with a EGCG treatment may have therapeutic potential in non-small cell lung cancers.

Granulocyte Macrophage-Colony Stimulating Factor (GM-CSF) Augments Acute Lung Injury via Its Neutrophil Priming Effects

Granulocyte macrophage-colony stimulating factor (GM-CSF) has immuno-stimulatory effects. We hypothesized that GM-CSF plays an important role both in lipopolysaccharide (LPS)- and hemorrhage-induced acute lung injury (ALI). We also postulated that GM-CSF augments LPS-induced inflammation by priming neutrophils. ALI was induced in GM-CSF-/- or control C57BL mice either by LPS injection or by hemorrhage. Lung inflammation (by lung expression for tumor necrosis factor-alpha (TNF-alpha), macrophage inflammatory protein-2 (MIP-2), interleukin-1beta (IL-1beta), interleukin- 6 (IL-6), and keratinocyte-derived chemokine) and lung injury (by myeloperoxidase and evans blue dye assay) were evaluated after ALI. Incremental doses of LPS (0, 1, 10, and 100 ng/mL) and GM-CSF (0, 1, 10, and 100 ng/mL) were added to bone marrow neutrophils. The expression of TNF-alpha, MIP-2, and IL-1beta was evaluated with enzyme linked immunosorbent assay. The mRNA expression of three cytokines, and the nuclear translocation of nuclear factor kappa B (NF kappa-B) were evaluated by reverse transcriptase-polymerase chain reaction and electropnoretic mobility shift assay, respectively. GM-CSF -/- mice showed decreased neutrophil infiltration, less leakage, and lower expression of cytokines in the lung after LPS or hemorrhage. GM-CSF augmented LPS-induced protein and mRNA expression of TNF-alpha, MIP-2 and IL-1beta, which was mediated by increased intra-nuclear translocation of NF-kappa B. GM-CSF plays an important role in high-dose LPS and hemorrhage-induced ALI, which appears to be mediated by its priming effect on neutrophils.

Causes of Fever in the ICU - A Prospective, Cohort Study / 대한구급학회지

BACKGROUND: Fever develops in 70% of ICU patients. In the present study, we tried to figure out causes of fever and the prognosis of febrile patients in the ICU in a prospective, cohort method. METHODS: From February to June 2007, patients admitted to medical ICU were daily screened and those who developed fever were enrolled. 237 consecutive admissions of 237 patients over a 5-month period were analyzed. Clinical parameters, including demographic data, underlying diseases, duration of ICU stay, causes of fever and final outcome were analyzed. RESULTS: Fever (core temperature > or =38.3degrees C) was present in 8% of admission, and it was caused by infective (84.2%) and non-infective processes (15.8%). Most fever occurred within first 5 days in the course of the admission (68.4%) and most lasted less than 5 days (57.9%). The median Acute Physiology and Chronic Health Evaluation (APACHE) III score at the time of fever was 43 (+/-19). Those with infectious fever had no significant differences in terms of severity of diseases in comparison with those with non-infectious cause of fever. The most common cause of infective fever was pneumonia (n=11). Prolonged fever (> or =5 days), all of which was caused by infection, occurred in 11 patients. Those with prolonged fever had higher mortality rate than short duration of fever (37.5% vs 0%, p<0.05). CONCLUSION: Infection, especially pneumonia is common cause of fever in the ICU. Prolonged fever is associated with high mortality rate.

A Case of Empyema and Mediastinitis by Non-typhi Salmonella / 결핵및호흡기질환

There are few reports of the pleuropulmonary involvement of a non-typhi Salmonella infection in immunocompromised patients with AIDS, malignancy, collagen vascular diseases, extended use of corticosteroids, sickle cell disease, or diabetes. We report a case of a non-immunocompromised patient who presented with concomitant empyema and mediastinitis due to Salmonella without a comorbid disease. A 26-year-old male patient, with a history of pneumonia 5 years earlier and having lived abroad for several years, presented chronic cough and febrile sensation. Pneumonia, empyema and mediastinitis were noted in a chest CT scan and Salmonella enteritidis and beta-hemolytic streptococcus were identified from a culture of the pleural fluid. Initially, he was treated with cefepime, metronidazole and clarithromycin. He was cured clinically and radiographically after an 8 week treatment with antibiotics. In conclusion, this report suggests that S. enteritidis can cause empyema and mediastinitis, albeit rarely.

Diagnostic Value of ADA Multiplied by Lymphocyte to Neutrophil Ratio in Tuberculous Pleurisy / 결핵및호흡기질환

BACKGROUND: Many diagnostic approaches for defining the definitive cause of pleurisy should be included due to the large variety of diseases resulting in pleural effusion. Although ADA is a useful diagnostic tool for making a differential diagnosis of pleural effusion, particularly for tuberculous pleural effusion, a definitive diagnostic cut-off value remains problematic in Korea. It was hypothesized that ADA multiplied by the Lymphocyte/Neutrophil ratio(L/N ratio) might be more powerful for making a differential diagnosis of pleural effusion. METHODS: One hundred and ninety patients, who underwent thoracentesis and treatment in Chung-Ang University Hospital from January, 2005 through to February 2006, were evaluated. The clinical characteristics, radiologic data and the examination of the pleural effusion were analyzed retrospectively. RESULTS: 1. Among the 190 patients, 59 patients (31.1%) were diagnosed with tuberculous pleurisy, 45 patients(23.7%) with parapneumonic effusion, 42 patients(22.1%) with malignant effusions, 36 patients(18.9%) with transudate, and 8 patients(4.2%) with empyema. One hundred and twenty one patients were found to have an ADA activity of 1 to 39 IU/L(63.7%). Twenty-nine were found to have an ADA activity of 40 to 75 IU/L(15.3%) and 40 were found to have an ADA activity of 75 IU/L or greater(21.0%). 2. Among the patients with tuberculous pleurisy, 5(8%), 18(30%) and 36 patients(60%) had an ADA activity ranging from 1 to 39 IU/L, 40 to 75 IU/L, and 75 IU/L or greater, respectively. In those with an ADA activitiy 40 to 75 IU/L, 18 patients(62%) had tuberculous pleurisy, 9(31%) had parapneumonic effusion and empyema, and 1(3.4%) had a malignant effusion. 3. In those with an ADA activity of 40 to 75 IU/L, there was no significant difference between tuberculous pleurisy and non-tuberculous pleural effusion(tuberculous pleurisy : 61.3 +/- 9.2 IU/L, non-tuberculous pleural effusion : 53.3+/-10.5 IU/L). 4. The mean L/N ratio of those with tuberculous pleurisy was 39.1 +/- 44.6, which was significantly higher than non- tuberculous pleural effusion patients (p<0.05). The mean ADA x L/N ratio of the tuberculous pleurisy patients was 2,445.7 +/- 2,818.5, which was significantly higher than the non-tuberculous pleural effusion patients (level p<0.05).5. ROC analysis showed that the ADA x L/N ratio had a higher diagnostic value than the ADA alone in the group with an ADA between 40-75 IU/L. CONCLUSION: The ADA multiplied by the lymphocyte-to-neutrophil ratio might provide a more definitive diagnosis of tuberculous pleurisy.

A Case of Tuberculous Lymphadenitis accompanying Papulonecrotic Tuberculid / 결핵및호흡기질환

Lymphadenitis is a common manifestation in tuberculous diseases. However, papulonecrotic tuberculid is an uncommon cutaneous manifestation, and is considered an allergic reaction against tuberculous bacilli in tuberculous lesions other than the lymph nodes. A wide great variety of cutaneous manifestations arise over a period of a few weeks - i.e., papules, necrosis, crusted and atrophic scars. We described a 27-year-old woman with right cervical tuberculous lymphadenitis and skin lesions involving her arms, legs and both fingers. Histopathologically, a leukocytoclastic vasculitis with V-shaped epidermal necrosis was observed in the upper and deep dermis, including the good response to anti-tuberculosis therapy support the diagnosis of papulonecrotic tuberculid.

Proinflammatory Effects of High Mobility Group B1 (HMGB1) Versus LPS and the Mechanism of IL-8 Promoter Stimulation by HMGB1 / 결핵및호흡기질환

BACKGROUND: High mobility group box 1 (HMGB1) is a novel, late mediator of inflammation. This study compared the pro-inflammatory effects of LPS and HMGB1. The transcriptional factors that play an important role in mediating the HMGB1-induced stimulation of IL-8 were also evaluated. METHODS: RAW264.7 cells were stimulated with either LPS (100 ng/ml) or HMGB1 (500 ng/ml). The TNF-alpha, MIP-2 and IL-1beta levels in the supernatant were evaluated by ELISA at 0, 2, 4, 8, 12 and 24h after stimulation. An acute lung injury was induced by an injection of LPS (5 mg/kg) or HMGB1 (2.5 mg/kg) into the peritoneum of the Balb/c mice. The lung cytokines and MPO activity were measured at 4h (for LPS) or 24h (for HMGB1) after the injection. The transcriptional factor binding sites for NF-IL6, NF-kappaB and AP-1 in the IL-8 promoter region were artificially mutated. Each mutant was ligated with pIL-6luc and transfected into the RAW264.7 cells. One hour after stimulation with HMGB1 (500 ng/ml), the cell lysate was analyzed for the luciferase activity. RESULTS: The expression of MIP-2, which peaked at 8h with LPS stimulation, increased sequentially until 24h after HMGB1 stimulation. An intraperitoneal injection of HMGB1, which induced a minimal increased in IL-1beta expression, provoked the accumulation of neutrophils the lung. A mutation of AP-1 as well as NF-kappaB in the IL-8 promoter region resulted in a lower luciferase activity after HMGB1 stimulation. CONCLUSION: The proinflammatory effects of HMGB1, particularly on IL-8, are mediated by both NF-kappaB and AP-1.

Microenvironments and Cellular Proliferation Affected by Oxygen Concentration in Non-Small Cell Lung Cancer Cell Line / 결핵및호흡기질환

BACKGROUND: Abnormal angiogenesis can induce hypoxia within a highly proliferating tumor mass, and these hypoxic conditions can in turn create clinical problems, such as resistance to chemotherapy. However, the mechanism by which hypoxia induces these changes has not yet been determined. Therefore, this study was conducted to determine how hypoxia induces changes in cell viability and extracellular microenvironments in an in vitro culture system using non-small cell lung cancer cells. METHODS: The non-small cell lung cancer cell line, A549 was cultured in DMEM or RPMI-1640 media that contained fetal bovine serum. A decrease in the oxygen tension of the media that contained the culture was then induced in a hypoxia microchamber using a CO2-N2 gas mixture. A gas analysis and an MTT assay were then conducted. RESULTS: (1) The decrease in oxygen tension was checked the anaerobic gas mixture for 30 min and then reoxygenation was induced by adding a 5% CO2-room air gas mixture to the chamber. (2) Purging with the anaerobic gas mixture was found to decrease the further oxygen tension of cell culture media. (3) The low oxygen tension resulted in a low pH, lactic acidosis and a decreased glucose concentration in the media. (4) The decrease in glucose concentration that was observed as a result of hypoxia was markedly different when different types of media were evaluated. (5) The decrease in oxygen tension inhibited proliferation of A549 cells. CONCLUSION: These data suggests that tumor hypoxia is associated with acidosis and hypoglycemia, which have been implicated in the development of resistance to chemotherapy and radiotherapy.