Recently the transcriptional up-regulation of human beta-defensin 2 (HBD-2) by lipopolysaccharide (LPS) was found to be associated with NF-kappaB binding site. Although the general mechanisms of NF-kappaB activation by LPS stimulation are well understood, less is known about the signal transduction pathway leading to LPS-induced NF-kappaB activation in human corneal epithelial (HCE) cells. The aim of this study was to investigate the intracellular signals involved in LPS-induced HBD-2 mRNA expression in HCE cells. Pretreatments of inhibitors for NF-kappaB, protein tyrosine kinase, p38 mitogen activated protein kinase (MAPK), and c-Jun N-terminal kinase (JNK) attenuated the LPS-induced NF-kappaB DNA binding activity and HBD-2 mRNA expression. Furthermore, pretreatments with inhibitors for protein kinase C (PKC), phosphatidylcholine-phospholipase C, phosphatidylinositol-phospholipase C, or phosphatidate phosphohydrolase prevented LPS-induced HBD-2 mRNA expression and HBD-2 prmoter-driven luciferase activity. However, the increased expression of HBD-2 mRNA and the increased DNA binding activity of NF-kappaB induced by LPS were not changed by the blockage of extracellular signal-regulated kinase (ERK) and of addition of antioxidants. Forskolin, a protein kinase A (PKA) agonist did not induce HBD-2 mRNA expression. These data demonstrate that LPS-induced HBD-2 mRNA expression via NF-kappaB is, at least in part, dependent on PKC, p38 MAPK, JNK, and protein tyrosine kinase status, but appears to be independent on PKA, ERK and ROS in HCE cells. Taken together, there may be more than one signaling pathways that lead to LPS-induced up-regulation of HBD-2 mRNA expression in HCE cells.
Antioxidants ; Binding Sites ; Colforsin ; Cyclic AMP-Dependent Protein Kinases ; DNA ; Epithelial Cells* ; Humans* ; JNK Mitogen-Activated Protein Kinases ; Luciferases ; NF-kappa B ; p38 Mitogen-Activated Protein Kinases ; Phosphatidate Phosphatase ; Phosphotransferases ; Protein Kinase C ; Protein Kinases ; Protein-Tyrosine Kinases ; RNA, Messenger* ; Signal Transduction ; Up-Regulation

Despite therapeutic advance, the prevalence of ischemic heart disease continues to increase. Recently, cell transplantation of stem cell has been proposed as a strategy for cardiac repair following myocardial damage. However, low differentiation efficiency into cardiomyocyte and poor cell viability associated with transplantation have limited the reparative capacity of these cell. In this study, we engineered P19 embryonal carcinoma cells using plasmid vector to overexpress the transcription factor MEF2c, Nkx2.5 involved in cardiomyogenesis. We investigated 1) formation of intercellular junction of P19 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. An P19 embryonal carcinoma cell line expressing GFP, MEF2c, Nkx2.5 was generated by gene transfection and clonal selection. Nkx2.5 overexpression induced connexin43 expression level decrease. Electron microscopy revealed myofibril organization and immunostaining with cTnT showed positive staining in P19-Nkx2.5, consistent with early stage cardiomyocyte. Connexin43 and N-cadherin was expressed between P19-MEF2c and cardiomyocyte, P19- Nkx2.5 and cardiomyocyte in co-culture. And beating rate of cardiomyocyte co-cultured with P19-Nkx2.5 increased much more than other group, even if P19-Nkx2.5 did not have synchronous contraction with cardiomyocyte. Additionally, P19-Nkx2.5 had a resistance against hypoxia. These result suggest that overexpression of Nkx2.5 induced differentiation of P19 into cardiomyocyte and would be electro-mechanical coupling with cardiomyocyte after transplantation. Futhermore, Nkx2.5 overexpression had protection potential to hypoxic injury. Therefore, P19 cell overexpressed Nkx2.5 would be promising cell source for further study of new therapy of myocardial disease and building up in vitro model.
Anoxia ; Cadherins ; Cardiomyopathies ; Cardiomyoplasty* ; Cell Survival ; Cell Transplantation ; Coculture Techniques ; Connexin 43 ; Embryonal Carcinoma Stem Cells ; Intercellular Junctions ; Microscopy, Electron ; Myocardial Ischemia ; Myocytes, Cardiac ; Myofibrils ; Plasmids ; Prevalence ; Stem Cells ; Transcription Factors* ; Transfection ; Transplants

Recently, new treatments for human heart disease such as ischemia, infarction, cardiomyopathy, coronary heart disease have been developed. transplantation various kinds of cells from skeletal muscle, endothelium, mesenchyme, hemopoietic tissue to injured area after infarction were challenged. It's so called 'Cell Transplantation'. This therapeutic strategy already adopted and got a good result in clinical trial. But several limitations are still remained, including ethics, donor cell numbers, side effects, therapeutic efficiency. In this research, we investigated the formation of intercellular junction and synchronous contraction between cardiomyocyte and H9c2 cell line in co-culture to establish experimental model in vitro for cell transplantation. For this purpose, two kinds of cells, primary cultured cardiomyocyte and H9c2 (cardiomyoblast cell line) were used. Cultured cardiomyocytes had repetitive contraction-relaxation pattern along longitudinal axis both in single and coculture. But their contractions were slower, less regular, less strong in co-culture than in cardiomyocyte culture only. H9c2 cells did not contracted actively themselves, but moved toward cardiomyocyte passively coincided with contraction. In contact region between two kinds of cells, there was no signal after immunocytochemical staining labeled with connexin43 (gap junction), desmoplakin (desmosome), N-cadherin (adherent junction) even though they had membrane contact. Moreover, F-actin and striation were less developed. These results suggested that co-culture system interfere with remodelling of contractile apparatus, intercellular junction formation as well as contraction-relaxation. Furthermore cardiomyocyte could not induce H9c2 cells differentiation into cardiomyocyte. Therefore, much more research would be essential for clinical application of cell transplantation and this study would be the basic source for further study of new therapy of myocardial disease and building up in vitro model.
Actins ; Axis, Cervical Vertebra ; Cadherins ; Cardiomyopathies ; Cell Count ; Cell Line* ; Cell Transplantation ; Coculture Techniques* ; Connexin 43 ; Coronary Disease ; Desmoplakins ; Endothelium ; Ethics ; Heart Diseases ; Humans ; Infarction ; Intercellular Junctions* ; Ischemia ; Membranes ; Mesoderm ; Models, Theoretical ; Muscle, Skeletal ; Myocytes, Cardiac* ; Tissue Donors ; Transplants

PURPOSE: To report two cases that showed clinical findings of recurrent keratoconus following penetrating keratoplasty. METHODS: A 39 year-old male and a 34 year-old female developed clinical signs of recurrent keratoconus in his left and her right eye 3 and 8 years after keratoplasty, respectively, and he underwent successful corneal regrafting. RESULTS: We confirmed recurrence of keratoconus histopathologically through the examination of the cornea of the recipient who underwent corneal regrafting and chromosomal study showed mosaicism, 47,XXY/46,XY. CONCLUSION: We report the histopathologic feature of one case of recurrent keratoconus for the first time in Korea.
Adult ; Cornea ; Corneal Transplantation ; Female ; Humans ; Keratoconus* ; Keratoplasty, Penetrating ; Korea ; Male ; Mosaicism ; Recurrence

Catechin is main component of polyphenol extracts from green tea, it is associated with prevention of hypertension and atherosclerosis, anti-diabetic effect, antioxidant, antitumor. The purpose of this research is to investigate the effect and its mechanism of green tea catechin on epithelial cancer cell lines in various concentrations and durations. For this study, epithelial cancer cell lines, A549 (lung cancer), EATC (Ehrlich-Lettre ascites tumor cell) were used. Inverted, light, confocal and electron microscopes were applied to find morphological changes. MTT assay, flowcytometric analysis, gel electrophoresis were used to compare severity of cellular damages to control after exposure to 1, 10, 100 and 500 microgram/ml catechin for 48 hours. In the A549 cells, after 1 microgram/ml and 10 microgram/ml catechin treatments, there was no notable changes. However, exposure to 100 microgram/ml catechin induced increase of cytoplasmic granules, destruction of lamellar body, inhibition of cell cycle, especially G0/G1. In the early phase of 500 microgram/ml catechin administration, decrease of cell population, severe destruction of lamellar bodies and mitochondria, derangement of cell cycle were shown. In the EATC, such as those effects occurred after exposure to lower concentration of catechin than in that of A549 cells. After exposure of 10 microgram/ml catechin, rounded-up cells and necrotic cells were found. Whereas, most of cells were under apoptotic changes-cytoplasmic condensation, nuclear fragmentation, cellular shrinkage, ladder pattern in the electrophoresis, when administrated 100 microgram/ml catechin. These results suggested that exposure of catechin induced severe cellular damage and growth inhibition in dose- and time-dependent manner. And we confirmed that these effects of catechin were involved with apoptosis, necrosis and cell cycle arrest and were quite different according to cancer type. Therefore, much more research would be demanded before clinical application of catechin to human cancer therapy and this study would be the basic source for further study of green tea.
Antioxidants ; Apoptosis ; Ascites ; Atherosclerosis ; Catechin* ; Cell Cycle ; Cell Cycle Checkpoints ; Cell Line* ; Cytoplasmic Granules ; Electrophoresis ; Humans ; Hypertension ; Mitochondria ; Necrosis ; Tea*

OBJECTIVE: In the management of spasticity, intramuscular neurolysis with small amount of dilute aqueous phenol has proved to be a useful measure. But, considerable problem has taken place in utilization of phenol. This study was attempted to compare the effect of phenol and alcohol for the peripheral nerve blocking in the management of spasticity. METHOD: Intraneural injection of 5% phenol, 50% alcohol and 90% alcohol solution carried out in each group of 10 rats. A total of 30 rat were injected and examined electrophysiologically before and after blocking the nerve (24 hour, 1 weeks, 2 weeks, 4 weeks, 8 weeks). The randomized one rat of each group was sacrificed for the histological examination of the sciatic nerve at every examined day. RESULTS: There was no difference of the distal latencies and amplitudes of compound muscle action potentials among the groups before injection. The latencies were prolonged at 24 hours post-injection and shortened at 1 week post-injection in all the groups. The amplitudes were markedly decreased at 24 hours post-injection and increased at 1 week post- injection and reached the pre-injection value at 8 week post-injection in all the groups. Histologic studies showed necrosis at 1 week post-injection and regeneration at 2 week post- injection in 50% and 90% ethanol groups. Phenol injection group showed necrosis at 4 week post-injection and regeneration after 8 weeks. CONCLUSION: Our preliminary experience with alcohol for peripheral nerve blocking with encouraging result has been described.
Action Potentials ; Animals ; Ethanol ; Muscle Spasticity ; Necrosis ; Nerve Block ; Peripheral Nerves ; Phenol* ; Rats* ; Regeneration ; Sciatic Nerve*

OBJECTIVE: To investigate the relationship between activities of daily living (ADL) dependence and psychological parameters in the adult hemiplegic stroke patients. METHODS: The subjects were total 122 stroke patients who underwent rehabilitation therapy. Patients were surveyed with questionnaires for anxiety, stress, and depression. Cognitive functions were assessed by mini-mental status examination and their ADL functions were evaluated by modified Barthel index. The correlation between ADL function and psychological parameters was statistically analyzed by Pearson's correlation test. RESULTS: Results were as follows: The average stress score of stroke patients was 76.7 which was relatively high. Fourty-seven percent of stroke patients suffered from depression, and it was more frequent in the left hemiplegics. As the result of the Pearson's correlation test, ADL function is closely correlated with intrapersonal stress level, but not with the level of anxiety or depression. The cognitive function has correlation with intrapersonal stress level, but not with depression. CONCLUSION: Understanding of the relationship between psychological status and ADL function in the stroke patients will be helpful to the physician for conducting efficient rehabilitation of these patients.
Activities of Daily Living* ; Adult ; Anxiety* ; Depression* ; Humans ; Surveys and Questionnaires ; Rehabilitation ; Stroke*

OBJECTIVE: To evaluate the effectiveness of nonsurgical treatment and to evaluate the usefulness of ultrasonographic assessment in postburn hypertrophic scar. METHODS: The subjects were twenty-seven burn patients with hypertrophic scar. Hypertrophic scars were treated by four different methods: triamcinolone acetonide injection (Group I), pressure garment application (Group II), combination treatment of triamcinolone acetonide and garment (Group III) and no treatment (Group IV). To assess the effectiveness of each treatment method, ultrasonographic measurement of scar thickness and punch biopsy of scar were done before and after treatment. RESULTS: After 4 weeks treatment, the scar thickness decreased by 14.9 9.4% in Group I, 4.7 8.4% in Group II and 20.5 13.2% in Group III. However the scar thickness increased by 10.0 13.5% in Group IV (no treatment). The pathologic findings didn't show significant change. CONCLUSION: The TA solution injection therapy and pressure garment therapy were effective methods for the treatment of the postburn hypertrophic scar, but the combinations of both therapies was more effective than each single therapy. The ultrasonography can be used as objective measure to assess effectiveness of therapy.
Biopsy ; Burns ; Cicatrix ; Cicatrix, Hypertrophic* ; Humans ; Triamcinolone Acetonide ; Ultrasonography

Medial femoral cutaneous nerve(MFCN), a sensory branch of the femoral nerve, supplies the skin over the anteromedial aspect of the thigh and knee. Posterior femoral cutaneous nerve(PFCN), comprised of fibers originating from the anterior and posterior divisions of the first three sacral segments, supplies the skin over the posterior aspect of the thigh. Forty nerves of twenty healthy adults, ages from 20 to 58, were tested. The onset and peak latencies of MFCN were 2.3+/-0.2 ms and 2.9+/-0.2 ms respectively. The baseline to peak amplitude was 6.5+/-2.3 V. The onset and peak latencies of PFCN were 2.4+/-0.2 ms and 2.9+/-0.2 ms respectively. The baseline to peak amplitude was 7.1+/-1.7 V.
Adult ; Equipment and Supplies ; Femoral Nerve ; Humans ; Knee ; Skin ; Thigh

Thirty-eight patients with a musculoskeletal chest wall syndrome were evaluated for the musculoskeletal findings of chest wall. All patients had the chest wall tenderness and the typical chest pain could be reproduced by the palpation. There was no significant difference in the diagnostic features of the pain for the onset, location, characteristics, duration, radiation, and area of references for chest pain among the different groups of the patients. However, a reproduction of pain by palpation and the pressure threshold difference between the lesion and control points by using pressure algometry was a reliable and specific diagnostic tool. Pressure threshold difference was correlated with numerical rating scale by the correlation coefficient 0.96. The common causes of the chest wall syndrome were the myofascial pain syndrome, chostochondritis, sternalis syndrome, rib-tip syndrome, xiphodynia in order. Six patients had chest wall disorders in conjunction with other associated intrathoracic condition. Thirty-two patients had an isolated chest wall syndrome. Chest wall syndrome should be considered in all patients with the chest pain, as its recognition could help the patient management.
Chest Pain ; Humans ; Myofascial Pain Syndromes ; Palpation ; Reproduction ; Thoracic Wall* ; Thorax*