Prevalence of Virulence Factors in Helicobacter pylori Isolated from Koreans and Proinflammatory Cytokine Gene Expression in Human Gastric Epithelial Cells Induced by H. pylori with Virulence Factors / 대한내과학회지

OBJECTIVES: CagA or cytotoxin-positive H. pylori may be associated with gastroduodenal diseases. However, controversies about this association also exist. Moreover, there could be geographic differences in the prevalence of virulence factors such as cagA or cytotoxin. In H. pylori infection, the gastric mucosa shows acute and chronic inflammation. However, the pathogenesis of such as an inflammation by H. pylori is not well elucidated. We performed this study 1) to determine prevalence of the genes of virulence factor such as cagA and cytotoxin in H. pylori, 2) to assess the correlation of their presence with clinical findings, and 3) to test whether the vacuolating cytotoxin of H. pylori could evoke proinflammatory cytokine gene expression in gastric epithelial cells. METHODS: 1) The prevalence of the cagA, vacA and adhesin genes in H. pylori strains isolated from Koreans was determined by PCR analysis. 2) H. pylori was cultured in Brucella broth containing 10% fetal bovine serum for 3 days using a shaker in a microaerophilic condition. Cytotoxin assay was performed by determining whether addition of the concentrated culture supernatants is able to cause vacuolization of HeLa cells. 3) After human gastric epithelial cells, Hs746T and AGS were incubated with the culture supernatants containing vacuolating cytotoxin, each RNAs were extracted from the gastric epithelial cells. And then various cytokine gene expression were assessed using RT-PCR. The expressed cytokine transcripts were quantified by RT-PCR and standard synthetic RNA. Among cytokines, IL-8 proteins were also measured by ELISA. RESULTS: 1) More than 95% of H. pylori isolates from Korean adults possessed cagA, vacA and adhesin genes. And 80.6% of H. pylori strains have expressed vacuolating cytotoxicity against HeLa cells within 24 hours. 2) There was no correlation between the virulence factors of H. pylori strains and clinical findings. 3) Cytotoxin-positive culture supernatants also caused vacuolization in gastric epithelial cells, both Hs746T and AGS. 4) Expression of mRNA for proinflammatory cytokines such as IL-1alpha: IL-8, MCP-1 and GM-CSF was much more upregulated by vacuolating cytotoxin-positive culture supernatants than cytotoxin-negative ones in both Hs746T and AGS cells. Number of molecules of the expressed IL-8 transcripts was parallel to the amounts of IL-8 protein secreted from gastric epithelial cells. CONCLUSION: These results suggest that virulence factors of H. pylori may not be factors determining disease entitiy in Korean patients infected with H. pylori. In addition, vacuolating cytotoxin secreted from H. pylori could give rise to vacuolization in gastric epithelial cells as well as induce proinflammatory cytokines from the cells.

The Effect of DA-6034, Eupatilin Derivatives of Flavonoids, on the Rats with Trinitrobenzene Sulfonic Acid (TNBS) - Induced Colitis / 대한내과학회지

OBJECTIVE: In traditional medicine, Artemisia capillaris has been used for treatment of chronic diarrhea. Previously we found Artemisia capillaris had an effect on rats with TNBS-induced colitis. Eupatilin, a kind of flavonoids, may be a probable effective component. To evaluate the effect of a eupatilin derivative compound DA-6034 on the rat with TNBS-induced colitis, we perfomed this study. METHODS: Colitis was induced with 1ml of 50 mg/ml TNBS mixed with 60 % ethanol (vol/vol) in Sprague- Dawley rats. From the next day, 1ml methylcellulose, 1 mg/kg prednisolone, 0.3 or 3 mg/kg of DA-6017 and DA-6034 were administered through rectum once daily for 2 weeks. At 2days, 1week, and 2weeks later, we evaluated the effect by gross damage score (0-10) and measured myeloperoxidase, PGE2, and LTB4 from the damaged mucosa. RESULTS: The mean gross damage scores of prednisolone and 3 mg/kg of DA-6034 groups were significantly lower than that of a placebo group at 2weeks (0.8, 0.9 vs. 4.0, p<0.05). Myeloperoxidase activities also seemed to be lower in those effective groups but were not statistically significant. LTB4 levels were lower in prednisolone and, 0.3 and 3 mg/kg of DA-6034 groups than in a placebo group at 2weeks (7.91, 7.23, and 7.13 vs. 13.90 ng/mg protein, p<0.05). PGE2 levels were decreased in prednisolone and 0.3 mg/kg of DA-6034 groups at 2days. DA-6017 showed no effects. CONCLUSIONS: Eupatilin derivative compound, DA- 6034 was effective in rats with TNBS-induced colitis. In that LTB4 level is lowered with some decrease of PGE2 level, this agent probably has an inhibitory effect on arachidonic acid metabolism.

Detection of Helicobacter DNA in the Bile from the Obstructed Bile Duct / 대한내과학회지

OBJECTIVE: Several newly recognized Helicobacter spp. such as H. hepaticus, H. bilis, H. cholecystus, H. rappini, H. pullorum, can cause persistent hepatitis, hepatoma, cholangiopancreatitis, and cholecystitis in animals. Recently some studies have been reported that Helicobacter DNA can be found in the bile from the patients with diseased bile duct, although its clinical significance is still unclear. The aim of this study is to investigate the existence, and character of Helicobacter in the bile from the obstructed bile duct, and the relationship with pH and the other bacteria found in the bile. METHODS: Twenty-eight bile samples (15 from bile duct cancer, 6 from pancreatic head cancer, 7 from bile duct stones) were obtained from the PTBD route. Bile pH measurement, and Helicobacter culture in microaerophil uric and anaerobic conditions were performed. The primers chosen for polymerase chain reaction (PCR) amplification for detection and characterization were ureA (411 bp) and cagA gene (298 bp), respectively. And primer of 16s rRNA for all known bacteria including Helicobacter was used, and the kinds of bacteria were identified by RFLP. RESULTS: Helicobacter DNA was detected in 39.3%. The bile pH was not related with presence of Helicobacter (7.83 +/-0.41 vs 7.78+/-0.48). The prevalence of cagA was 35.7%, and 16s rRNA was found in 46.4%. The specific 16s rRNA band for Helicobacter was observed in 14.3%. All the culture were not successful. CONCLUSION: Although the Helicobacter spp. were not cultured, Helicobacter exists obviously in the bile from the diseased bile duct, and coexist with other bacteria. These results should stimulate studies to ascertain whether these Helicobacter play a role in the pathogenesis of bile duct diseases in human.

Evaluation of the Left Atrial Size and Function in Addition to Analysis of the Mitral and Pulmonary Venous Flow Velocity in the Estimation of Left Ventricular Filling Pressures

BACKGROUND: Mitral and pulmonary venous(PV) flow velocity variables are being used for the indirect evaluation of left ventricular(LV) diastolic function. However, these flow velocities are influenced by age, loading conditions and other factors. This study was designed to evaluate usefulness of left atrial size and function in addition to the relation of mitral and PV flow velocity variables in the estimation of LV filling pressures. METHODS: Mitral and PV flow velocity variables. left artial size and function were assessed just before a cardiac catherization in 31 patients. According to the LV filling pressures, patients were divided into two subgroups and echocardiographic variables were compared. RESULTS: 1) LV end-diastolic pressure was related to the duration of reverse flow in the PV at atrial contraction(r=0.58) and difference in mitral and PV flow velocity duration at atrial contraction(r=0.54), and the similar findings were observed in other left ventricular filling pressures. 2) Left atrial size and volumes were greater in the subgroup of abnormal LV filling pressures(P < 0.05), but left atrial ejection fraction was not different between subgroups. CONCLUSION: In addition to variables of the mitral and PV flow velocities, left atrial size and volume may provide an additive value in the estimation of left ventricular filling pressures.

Bilateral Sequential Bronchopulmonary Lavage for Pulmonary Alveolar Proteinosis / 대한마취과학회지

Pulmonary alveolar proteinosis is a rare disease of unknown etiology characterized by the accumulation of surfactant like phospholipids and proteinaceous material in the alveolar space. The removal of lipoproteinaceous material from the alveolar can be the only means of effectively treating the progressive hypoxemia in pulmonary alveolar proteinosis. The current technique of the lavage follows: The procedure can be accomplished by the use of Carlens tube, with lavaging of one lung while the other is ventilated, the ventilated lung is lavaged in a similar manner two or three days later. In this case, we performed sequential bilateral pulmonary lavage. We report a case of broncho-pulmonary lavage in which changes of oxygenation and hemodynamics were observed. We conclude that the procedure can be performed by careful monitoring such as continuous monitoring of S v O2, SpO2, effective static comliance.