Cellular cardiomyoplasty has recently emerged as a potential new treatment of ischemic heart disease. Combining cellular cardiomyoplasty with gene therapy using myogenic transcription factor might facilitate myocardial regeneration. In this study, we engineered H9c2, L6 using plasmid vector to overexpress the transcription factor MEF2c, Nkx2.5 involved in cardiomyogenesis. We investigated 1) formation of intercellular junction in mono-culture and co-culture with cardiomyocyte for functional and structural synchronous contraction after transplantation, 2) differentiation into cardiomyocyte, 3) resistance to hypoxic condition. Each cell overexpressing MEF2 and Nkx2.5 was generated by gene transfection and clonal selection. CO-culture was performed that each cell line added over cultured cardiomyocyte. H9c2-MEF2c and H9c2-Nkx2.5 became long, spindle shape like cardiomyocyte. Troponin T, cardiac specific marker, was found spot-like pattern in H9c2-Nkx2.5. However, co-culture with cardiomyocyte did not induce differentiation all kinds of cells into cardiomyocyte. Connexin43, which is gap junction marker was increased in H9c2-MEF2c, H9c2-Nkx2.5, L6-MEF2c and L6-Nkx2.5. Especially, co-culture with cardiomyocyte resulted in elevation of connexin43 levels more than monoculture. Ultrastructurally, formations of gap junction and desmosome were found apparently in L6-Nkx2.5. Long-standing, strong, regular and more frequent contraction were observed in cardiomyocyte co-cultured with H9c2-MEF2c, H9c2-Nkx2.5, L6-MEF2c, L6-Nkx2.5, respectively. Neverthless, any cell did not have active contraction itself, but passive movement except cardiomyocyte. H9c2-MEF2c, L6-MEF2c and L6-Nkx2.5 had resistance to hypoxia compared with other groups. These results suggested that co-culture and overexpressions of MEF2c and Nkx2.5 induced differentiation into cardiomyocyte and played an important role on intercellular junction formation and hypoxic resistance. This would be a promising source of cellular cardiomyoplasty. Therefore, much more research would be essential for clinical application of cellular cardiomyoplasty and this study would be a basic source for further study of MEF2c and Nkx2.5 in cellular cardiomyoplasty.
Animals ; Anoxia ; Cardiomyoplasty ; Cell Line ; Cell Transplantation ; Coculture Techniques* ; Connexin 43 ; Desmosomes ; Gap Junctions ; Genetic Therapy ; Intercellular Junctions ; Myoblasts ; Myoblasts, Cardiac* ; Myocardial Ischemia ; Myocytes, Cardiac* ; Plasmids ; Rats* ; Regeneration ; Transcription Factors ; Transfection ; Troponin T

BACKGROUND AND OBJECTIVE: Even though they are identified on the basis of their induction by elevated temperatures, heat shock proteins are induced by various stimuli. Such inducers include viral infection, exposure to ethanol, sodium arsenite, steroid hormones and heavy metals. They become involved under pathological conditions, such as anoxia/ischemia, reperfusion injury and oxidative stress in the heart. In this research, the cellular damages and changes of Hsp25 and alphaB crytallin were investigated under oxidative stress induced H2O2 treatment in a cultured cardiomyoblast cell line, H9c2. MATERIALS AND METHODS: The morphological changes of the H9c2, and the distributions of Hsp25 and alphaB crytallin, were examined with phase-contrast, confocal and electron microscopy. The levels of Hsp25 and alphaB crytallin expressions within the cells were also checked. RESULTS: After exposure to a low concentration (10 micrometer) of H2O2, the amounts of Hsp25 and alphaB crytallin were increased more than in the control and heat shock groups. They were particularly localized in the perinuclear region and along the actin filament. When exposed to a high concentration (more than 100 micrometer) of H2O2, many cells developed apoptotic features, such as vacuolization, blebbing and apoptotic body formations. The amounts of Hsp25 and alphaB crytallin were decreased more than in the control group. However, a weak reaction in the perinuclear region and along the actin filaments still remained when observed under confocal microscopy. CONCLUSION: These results suggest that the elevation of the expressions of Hsp25 and alphaB crytallin were dependent on the concentration of H2O2. Additionally, the redistribution of these small Hsps is closely associated with the actin filaments, as well as the nuclei. Therefore, much more research will be essential for the elucidation of roles of Hsp25 and alphaB crytallin in oxidative stress, such as ischemia or ischemic-reperfusion injury.
Actin Cytoskeleton ; Actins* ; Blister ; Cell Line* ; Ethanol ; Heart ; Heat-Shock Proteins* ; Hot Temperature* ; Ischemia ; Metals, Heavy ; Microscopy, Confocal ; Microscopy, Electron ; Oxidative Stress ; Reperfusion Injury ; Shock ; Sodium

Diatomology has provided important evidences for diagnosis of drowning in forensic practice. However, conflicting opinions about the reliability of diatom test still exist. As a part of efforts to end this controversy, it is very important to secure the diatom profile in the water sample of drowning site. Our results obtained in this study show the characteristics of temporal and spatial distribution of diatom collected from the main drowning sites of Han river in Korea. During spring, 111 taxa (98 species and 13 varieties) were identified from 31 genera, and 87 taxa (77 species and 10 varieties) were identified from 22 genera during summer. At the level of genus Achnanthes, Cyclotella, Cymbella, Fragilaria, Navicula, Nitzschia, and Stephanodiscus represented the highest numbers, and were the most widely occurred during two seasons. The centric diatoms, Stephanodiscus hantzschii f. tenuis and Cyclotella atomus were typical of spring and summer, respectively, and identified as dominant indicator species. The commonly occurred species, including dominant species in the seasonal cluster analysis influenced the formation of site - specific groups. As a result, our results will be able to be used not only as the comparison data of diatom found in the drowning victim's tissue but also a reference data to rule out the possibility of contamination.
Diatoms ; Drowning ; Korea ; Rivers ; Seasons ; Water

BACKGROUND: Non-small cell carcinoma (NSCC) has become the leading cause of cancer related death around the world. However, its prognostic factors remain poorly defined. Galectin-3 is an apoptosis related protein and its relationship with various cancers is presently the subject of research. This study was performed to investigate galectin-3 expression in NSCC and its value as a prognostic factor. METHODS: We examined the expression of galectin-3 and bcl-2 in surgically resected, lung NSCC, including 61 squamous cell carcinomas and 41 adenocarcinomas. PCNA staining was also performed. RESULTS: Each type of carcinoma showed cytoplasmic positivity in 18 (30.0%) and 25 (61.0%) cases, respectively. Squamous cell carcinoma showed increased galectin-3 expression in better differentiated tumors, whereas adenocarcinoma didn't show any relationship with the degree of differentiation. The cytoplasmic positivity of galectin-3 in both types of carcinoma was associated with poor prognosis. Bcl-2 expression didn't show any significant relationship with overall survival. Galectin-3 and bcl-2 expressions were positively correlated. However, co-expression of both proteins was not related to prognosis. CONCLUSIONS: These results indicate that galectin-3 expression in NSCC warrants attention as a possible prognostic factor.
Adenocarcinoma ; Apoptosis ; Carcinoma, Non-Small-Cell Lung ; Carcinoma, Squamous Cell ; Cytoplasm ; Galectin 3* ; Lung ; Prognosis ; Proliferating Cell Nuclear Antigen

Mammary carcinoma with osteoclast-like giant cells is an unusual neoplasm characterized by giant cells, mononuclear stromal cells, and hemorrhage accompanying a low grade carcinoma. We present the cytological findings in a case of invasive ductal carcinoma with osteoclast-like giant cells that was initially confused with a fibroadenoma, due to its well-demarcated and soft mass and the young age of the patient. A 28-year-old female presented with a 4.5 cm, well demarcated, soft and nontender mass in the right breast. Fine needle aspiration cytology (FNAC) showed a combination of low grade malignant epithelial cell clusters and osteoclast-like giant cells. The atypical epithelial cells were present in cohesive sheets and clusters. Osteoclast-like giant cells and bland-looking mononuclear cells were scattered. An histological examination revealed the presence of an invasive ductal carcinoma with osteoclast-like giant cells. We report here the cytological findings of this rare carcinoma in a very young woman. The minimal atypia of the epithelial cells and its soft consistency may lead to a false negative diagnosis in a young woman. The recognition that osteoclastlike giant cells are rarely present in a low grade carcinoma, but not in benign lesion, can assist the physician in making a correct diagnosis.
Adult ; Biopsy, Fine-Needle ; Breast ; Carcinoma, Ductal* ; Diagnosis ; Epithelial Cells ; Female ; Fibroadenoma ; Giant Cells* ; Hemorrhage ; Humans ; Stromal Cells

Although isolated contralateral tongue deviation following unilateral cortical infarction was occasionally reported, the unilateral lesion usually produces no significant deficit of tongue motility considering bilateral supranuclear innervation of the hypoglossal nerve. We observed a patient with obvious tongue paralysis, including intrinsic muscles, caused by ischemic stroke involving the motor area of the tongue in the primary motor cortex.

PURPOSE: The purpose of this study is to identify useful clinicopathologic factors for the prediction of lymph node metastasis in submucosally invasive colorectal carcinoma. METHODS: A total of fifty-four cases of colorectal carcinomas with submucosal invasion were included. The patients underwent curative resection with en bloc lymph node dissection. Clinical features such as age, gender, tumor size and tumor location were reviewed. Histopathologic examinations for tumor growth type, differentiation, depth of tumor invasion, lymphovascular invasion, neural invasion, tumor budding and peritumoral inflammation were performed. The expression of E-cadherin, beta-catenin, Smad4, p53 and Ki-67 were examined by immunohistochemistry. The correlation between the clinicopathologic factors and lymph node metastasis was evaluated. RESULTS: From the 54 patients with submucosally invasivecolorectal carcinoma, lymph node metastasis was identified in 13 cases (24.1%). The incidence of lymph node metastasis was significantly higher in cases positive for lymphovascular invasion (55.6% vs. 17.8%, P=0.028) and positive for tumor budding (47.4% vs. 11.45%, P=0.006). Cases negative for Smad4 had a higher tendency for incidence of lymph node metastasis (28.6% vs. 15.8%, P=0.341). Other clinicopathologic and immunohistochemical features were irrelevant to the lymph node status. In multivariate analysis, only tumor budding was an independent predictor of lymph node metastasis (P=0.051). CONCLUSION: Lymphovascular invasion and tumor budding were predictive factors of lymph node metastasis in submucosally invasive colorectal carcinoma. The incidence of lymph node metastasis of submucosally invasive colorectal carcinoma was not low. Careful selection for avoiding surgery in submuocally invasive colorectal carcinoma should be considered.
beta Catenin ; Cadherins ; Colorectal Neoplasms ; Humans ; Immunohistochemistry ; Incidence ; Inflammation ; Lymph Node Excision ; Lymph Nodes ; Multivariate Analysis ; Neoplasm Metastasis

OBJECTIVE: To investigate the presence of cranial neuropathy in patients with platinum-analogue chemotherapy using electrodiagnostic evaluations. METHODS: Thirty-nine patients whose chemotherapy was completed within a month and 40 control subjects were enrolled in the study. Electrodiagnostic evaluation was performed using sensory and motor nerve conduction studies and blink reflex studies, in addition to the two-point discrimination test. RESULTS: The chemotherapy group had significantly longer latencies of bilateral R1 responses (left p<0.001; right p<0.001) and greater distance in two-point discrimination (p<0.001) compared to the control group. In the subgroup with peripheral polyneuropathy, the left R1 (p=0.01), both R2i (left p=0.02; right p=0.03) and the left R2c (p=0.02) were prolonged relative to those without the polyneuropathy, and both R1 (left p<0.001; right p<0.001), R2i (left p=0.01; right p=0.03), and the left R2c (p=0.01) were prolonged relative to the controls. On the other hand, the subgroup without the polyneuropathy showed only prolongation of both R1 (left p=0.006; right p<0.001) relative to the controls. CONCLUSION: In the present study, comparison of blink reflex and two-point discrimination showed the likelihood of subclinical cranial neuropathy following platinum-analogue chemotherapy. Cranial neuropathy caused by platinum agents was more profound in patients with peripheral polyneuropathy and may be dependent on the cumulative dose of the drug. The blink reflex may be of value in detecting subclinical cranial neuropathy in patients undergoing platinum-analogue chemotherapy.
Blinking* ; Cranial Nerve Diseases ; Cranial Nerves ; Discrimination (Psychology) ; Drug Therapy* ; Hand ; Humans ; Neural Conduction ; Platinum ; Platinum Compounds ; Polyneuropathies

BACKGROUND: Topoisomerase II (TOPO II) is an enzyme that separates intertwined chromosomes during DNA synthesis by transiently breaking and joining DNA strands. The level of TOP II is one of the determinants of cellular sensitivity to anti-tumor drugs in non-Hodgkin's lymphoma patients. The alpha form of TOPO II has been recently used as a marker of cellular proliferation. High levels of TOPO IIalpha are expressed in aggressive and proliferative tumors. METHODS: This study was designed to evaluate the relationship between TOPO IIalpha expression and clinicopathological parameters including age, gender, the serum LDH level, the serum beta2-microglobulin level and stage, or expressions, of Ki-67, p53 and p27, in non-Hodgkin's lymphoma. We analyzed forty-one biopsied tissue specimens from patients with non-Hodgkin's lymphoma. RESULTS: The expression of TOPO IIalpha increased with the clinical stage and it was correlated with Ki-67 and p53 expressions. However, TOPO IIalpha expression did not have any significant correlation with age, gender, the serum LDH level, the serum 2-microglobulin level and the p27 expression. CONCLUSIONS: TOPO IIalpha expression is a useful marker of cellular proliferation and it may serve as a prognostic factor of a tumor's progression and aggressiveness in non-Hodgkin's lymphomas.
Cell Proliferation ; DNA Topoisomerases, Type I* ; DNA Topoisomerases, Type II ; DNA* ; Humans ; Ki-67 Antigen ; Lymphoma, Non-Hodgkin*

BACKGROUND: The aim of this study was to clarify the incidence and role of microsatellite instability (MSI) in sporadic ovarian epithelial cancers (OEC). We investigated the MSI status and mismatch repair (MMR) protein expression in OEC. METHODS: MSI was examined by fluorescence- based polymerase chain reaction using five NCI panel markers (BAT25, BAT26, D2S123, D5S346 and D17S250) in 46 cases of OEC. Immunohistochemistry (IHC) for hMLH1 and hMSH2 was performed. RESULTS: Seven cases (15.2%) exhibited high-frequency MSI (MSIH), one exhibited low-frequency MSI (MSI-L), and the remaining 38 demonstrated microsatellite stability (MSS). MSI-H in OEC was not associated with histologic grade, FIGO stage, tumor size, mitoses or histologic type. Loss of expression of either hMLH1 or hMSH2 was observed in 4 of the 7 (59.3%) MSI-H cases, whereas 4 of the 39 (10.3%) MSI-L or MSS tumors revealed loss of expression of MMR proteins. The sensitivity and specificity of immunohistochemistry for hMLH1 and hMSH2 were 57.1% and 89.7%. CONCLUSIONS: Our data suggest that a genetic defect in the MMR system might play a role in the carcinogenesis of a minor subset of sporadic OEC however, immunohistochemical testing for hMLH1 and hMSH2 cannot accurately determine microsatellite instability status in OEC.
Carcinogenesis ; DNA Mismatch Repair ; Immunohistochemistry ; Incidence ; Microsatellite Instability* ; Microsatellite Repeats* ; Mitosis ; Ovarian Neoplasms ; Polymerase Chain Reaction ; Sensitivity and Specificity