To obtain an adequate quality and quantity of DNA from formalin-fixed and paraffin-embedded tissue, six different DNA extraction methods were compared. Four methods used deparaffinization by xylene followed by proteinase K digestion and phenol-chloroform extraction. The temperature of the different steps was changed to obtain higher yields and improved quality of extracted DNA. The remaining two methods used microwave heating for deparaffinization. The best DNA extraction method consisted of deparaffinization by microwave irradiation, protein digestion with proteinase K at 48°C overnight, and no further purification steps. By this method, the highest DNA yield was obtained and the amplification of a 989-base pair β-globin gene fragment was achieved. Furthermore, DNA extracted by means of this procedure from five gastric carcinomas was successfully used for single strand conformation polymorphism and direct sequencing assays of the β-catenin gene. Because the microwave-based DNA extraction method presented here is simple, has a lower contamination risk, and results in a higher yield of DNA compared with the ordinary organic chemical reagent–based extraction method, it is considered applicable to various clinical and basic fields.

Background The diagnosis of acute coronary syndrome (ACS), especially non-ST-elevation myocardial infarction and unstable angina in the emergency department (ED) still remains a challenge. Multislice computed tomography (MSCT) allows assessment of not only coronary artery stenoses and occlusions, but also assessment of coronary artery plaques and myocardial perfusion status. Methods and Results MSCT was performed in 31 patients who were admitted to the ED because of chest pain persisting at least 30 min and non-diagnostic ECG changes and normal serum enzyme concentrations. Using MSCT, ACS was defined by coronary artery stenosis ≥75% accompanied by computed tomography (CT)-low-density plaques, and/or by the presence of myocardial perfusion defects. ACS was confirmed by coronary stenosis ≥75% by coronary angiography and/or subsequent elevation of troponin I concentration. In total, 22 patients were diagnosed as having ACS. MSCT detected stenoses with CT-low-density plaques in 21 and non-transmural myocardial perfusion defect in 3 patients. There was 1 false-positive and 1 false-negative result. The sensitivity and specificity of MSCT to identify ACS was 95.5% and 88.9%, respectively. Conclusion MSCT provides diagnostic operating characteristics suitable for triage of patients with ACS in the ED. (Circ J 2005; 69: 1047 - 1051)

Background In the present study, multislice spiral computed tomography (MSCT), which allows non-invasive assessment of coronary artery plaque, was used to compare the CT density of plaque between patients with acute coronary syndrome (ACS) and those with stable angina (SA). Methods and Results MSCT was performed in 20 patients with ACS (17 with acute myocardial infarction, 3 with unstable angina) and 22 patients with SA. The presence of the plaque was defined on the basis of multiplanar reformation and axial images. At least 4 regions of interest were then placed within the plaque and the minimum CT density was measured and expressed as Hounsfield units (HU). The number of plaques did not differ between the 2 groups, but the minimum CT density was significantly lower in patients with ACS (25±15 HU) than in those with SA (71±16 HU, range 46-101 HU, p<0.001). Similarly, the minimum plaque density was significantly lower in the culprit coronary segment (26±16 HU) than in the non-culprit segment (48±17 HU) in 15 ACS patients with multiple plaques. Conclusion MSCT can potentially differentiate vulnerable from stable plaque in patients with coronary artery disease, although long-term, prospective analysis is needed to establish the conclusion. (Circ J 2004; 68: 840 - 844)

To investigate the relationships between the expression of MUC5B and clinicopathological parameters, the expression of MUC5B was immunohistochemically studied. MUC5B expression was observed in 129 of 198 (65.2%) adenocarcinomas and in 4 of 49 (8.2%) squamous cell carcinomas (P < 0.00001). MUC5B expression was significantly associated with poorer differentiation (P = 0.0303), higher pathological TNM stage (p = 0.0153) and poorer prognosis of adenocarcinoma patients (P = 0.0017). Multivariable analysis with Cox proportional hazards models confirmed that MUC5B expression increased the hazard of death after adjusting for other clinicopathological factors (HR = 2.66; 95%CI, 1.26–5.61). We also immunohistochemically evaluated TTF-1 expression and found that the combination of MUC5B with TTF-1 is a useful marker for adenocarcinomas. The diagnostic accuracies of TTF-1 and MUC5B for adenocarcinoma were 83.8% and 70.4%, respectively. The accuracy increased to 94.3% when the two factors were combined. In survival analysis, the MUC5B(High)/TTF-1(−) group was significantly associated with a poorer outcome compared with the MUC5B(Low)/TTF-1(+) group (p < 0.0001). The present study suggested that the combination of MUC5B and TTF-1 expression is useful for discriminating adenocarcinomas from squamous cell carcinomas, yielding prognostic significance in patients with lung adenocarcinoma.

Myosin-9 (MYH9) belongs to the myosin superfamily of actin-binding motor protein. Recently, MYH9 has been thought to be associated with cancer cell migration, invasion, and metastasis. The aims of this study were to immunohistochemically examine MYH9 expression in surgically resected non-small cell lung cancer (NSCLC), and evaluate its correlations with clinicopathological parameters and the prognosis of patients. MYH9 expression was immunohistochemically studied in 266 consecutive resected NSCLCs, and its associations with clinicopathological parameters were evaluated. Kaplan-Meier survival analysis and Cox proportional hazards models were used to estimate the effect of MYH9 expression on survival. MYH9 expression was detected in 102 of 266 (38.3%) NSCLCs. MYH9 expression was significantly correlated with the adenocarcinoma histology (P = 0.014), poorer differentiation ((P = 0.033), intratumoral vascular invasion and lymphatic invasion ((P = 0.013 and P = 0.045 respectively), and a poorer prognosis ((P = 0.032). In addition, multivariable analysis revealed that MYH9 expression independently predicted a poorer survival (HR, 2.15; 95%CI, 1.17-3.92; (P = 0.01). The present study revealed that MYH9 is expressed in a subset of NSCLC with a more malignant nature, and its expression is an indicator of a poorer survival probability.