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

Recently, tibial shaft fractures become more severe because of high speed traffic accidents. The open tibial fracture is a difficult one to treat because of its common complications such as infection, soft tissue defect, delayed union and non-union. We studied 37 cases of open tibial fractures which were treated with external fixation. The results are as follows: 1. The average patient age was 29.3 years and male predominated by a ratio of 4.3: l. 2. The traffic accidents were the most common cause of injury. 3. Most cases(67.6%) were type III fracture(by classification of Gustilo and Anderson). 4. Hoffmann external fixator gave rigid fixation and good wound access. 5. There were 28 cases(75.7%) which needed secondary soft tissue reconstruction. 6. The infection rate was 21.5%(8 cases). These all healed with conservative management. There were 5 cases of pin tract infection which healed without developing osteitis. 7. The average union time was 28.7 weeks. In type III fracture there was marked delay in union (average 32.1 weeks). 8. The rate of development of delayed or non-union was 35.1%. In these cases, bony union was obtained with cancellous bone graft.
Accidents, Traffic ; Classification ; External Fixators ; Humans ; Male ; Osteitis ; Soft Tissue Infections ; Tibia ; Tibial Fractures ; Transplants ; Wounds and Injuries

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

A heterogenic group of staphylococcal exotoxins, including staphylococcal superantigenic toxins, enterotoxin (SE), toxic shock syndrome toxin-1 (TSST-1), and coagulase are the most important virulence factors of Staphylococcus aureus. We analyzed the prevalence of genes encoding five enterotoxins and TSST-1 in S. aureus isolated from clinical ear discharges. The genes were identified by multiplex PCR and we compared the results to references of coagulase serotypes. In 102 isolates of S. aureus, 44 of them were methicillin-resistant S. aureus (MRSA) and the others were methicillin-susceptible S. aureus (MSSA). Among both types of S. aureus, 33 strains were positive for sea, 2 for seb, 23 for sec, 26 for see, and 26 for tst. Overall, 59 (57.8%) isolates were positive for one or more superantigenic toxin genes. From these, 71.2% (42/59) strains harbored more than one toxin gene in different combinations. The major combinations of genes were sea and see, and sec and tst. The degree of possession of superantigenic toxic genes was similar in both MRSA and MSSA isolates (56.8% vs 58.6%, respectively), yet significant differences in toxin gene profiles and coagulase serotypes between two isolates were detected. All of 13 positive strains for sec and tst were MRSA and belonged to coagulase serotype II. On the other hand, 80.0% of 20 positive strains for sea and see were MSSA with coagulase serotype IV and VII, whereas 20.0% of them were MRSA with coagulase serotype IV. This data indicates that the profile of superantigenic toxin genes correlates to coagulase serotype and methicillin resistance in S. aureus isolates.
Bacterial Toxins ; Coagulase ; Ear ; Enterotoxins ; Exotoxins ; Hand ; Methicillin Resistance ; Methicillin-Resistant Staphylococcus aureus ; Multiplex Polymerase Chain Reaction ; Prevalence ; Shock, Septic ; Staphylococcus ; Staphylococcus aureus ; Superantigens ; Virulence Factors

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: Postoperative pancreatic fistula (POPF) is the most troublesome complication after pancreaticojejunostomy (PJ).This study aimed to compare the short-term outcomes of 2 different methods of duct-to-mucosa PJ; out-layer continuous suture anastomosis (OCA) and the modified Blumgart method (mBM). Methods: This retrospective cohort study enrolled patients who underwent curative-intent, open PD between 2015 and 2020. In mBM, 2 transpancreatic U-sutures were performed between the pancreatic margin and jejunum, with reinforced sutures in the central region. Patient demographics, diagnosis, intraoperative factors, postoperative complications, and POPF defined by the International Study Group on Pancreatic Fistula were investigated. Clinically relevant POPF (CR-POPF) included grades B and C POPF. Results: A total of 184 patients underwent OCA, and 96 patients underwent mBM. The mBM group had more patients who underwent neoadjuvant therapy. The fistula risk scores were comparable between the 2 groups. Both groups showed no significant differences in CR-POPF and overall surgical complication rates. The total operation time was comparable, although the operation time for PJ was shorter in mBM. Conclusion No significant differences were observed in the postoperative outcomes between each group; the operation time for PJ in mBM was shorter. Therefore, mBM may be considered for utilization in duct-to-mucosa PJ.

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

The authors report a successful thrombectomy using the AngioJet Rheolytic Thrombectomy System (AngioJet) and an embolic protection device in a patient with femorofemoral bypass graft occlusion. Lower extremity CT angiograms showed occlusion in the left-to-right femorofemoral bypass graft. A rheolytic thrombectomy using the AngioJet and balloon angioplasty restored blood flow to the right lower extremity, and distal embolization may be effectively prevented by placing an embolic protection device within the right superficial femoral artery during the procedure.