The purpose of current experiment is the generation of insulin-producing human mesenchymal stem cells as therapeutic source for the cure of type 1 diabetes. Type 1 diabetes is generally caused by insulin deficiency accompanied by the destruction of islet beta-cells. In various trials for the treatment of type 1 diabetes, cell-based gene therapy using stem cells is considered as one of the most useful candidate for the treatment. In this experiment, human mesenchymal stem cells were transduced with AAV which is containing furin-cleavable human preproinsulin gene to generate insulin-producing cells as surrogate beta-cells for the type 1 diabetes therapy. In the rAAV production procedure, rAAV was generated by transfection of AD293 cells. Human mesenchymal stems cells were transduced using rAAV with a various multiplicity of infection. Transduction of recombinant AAV was also tested using beta-galactosidse expression. Cell viability was determined by using MTT assay to evaluate the toxicity of the transduction procedure. Expression and production of Insulin were tested using reverse transcriptase-polymerase chain reaction and immunocytochemistry. Secretion of human insulin and C-peptide from the cells was assayed using enzyme-linked immunosorbent assay. Production of insulin and C-peptide from the test group represented a higher increase compared to the control group. In this study, we examined generation of insulin-producing cells from mesenchymal stem cells by genetic engineering for diabetes therapy. This work might be valuable to the field of tissue engineering for diabetes treatment.

This study investigated that whether a 2 mT, 60 Hz, sinusoidal electromagnetic field (EMF) alters the structure and function of cells. This research compared the effects of EMF on four kinds of cell lines: hFOB 1.19 (fetal osteoblast), T/G HA-VSMC (aortic vascular smooth muscle cell), RPMI 7666 (B lymphoblast), and HCN-2 (cortical neuronal cell). Over 14 days, cells were exposed to EMF for 1, 3, or 6 hours per day (hrs/d). The results pointed to a cell type-specific reaction to EMF exposure. In addition, the cellular responses were dependent on duration of EMF exposure. In the present study, cell proliferation was the trait most sensitive to EMF. EMF treatment promoted growth of hFOB 1.19 and HCN-2 compared with control cells at 7 and 14 days of incubation. When the exposure time was 3 hrs/d, EMF enhanced the proliferation of RPMI 7666 but inhibited that of T/G HA- VSMC. On the other hand, the effects of EMF on cell cycle distribution, cell differentiation, and actin distribution were unclear. Furthermore, we hardly found any correlation between EMF exposure and gap junctional intercellular communication in hFOB 1.19. This study revealed that EMF might serve as a potential tool for manipulating cell proliferation.
Signal Transduction ; Microfilaments/radiation effects ; Humans ; Gap Junctions/metabolism/radiation effects ; *Electromagnetic Fields ; Cell Proliferation/radiation effects ; Cell Physiology/*radiation effects ; Cell Line ; Cell Differentiation/radiation effects ; Cell Cycle/radiation effects

This study investigated that whether a 2 mT, 60 Hz, sinusoidal electromagnetic field (EMF) alters the structure and function of cells. This research compared the effects of EMF on four kinds of cell lines: hFOB 1.19 (fetal osteoblast), T/G HA-VSMC (aortic vascular smooth muscle cell), RPMI 7666 (B lymphoblast), and HCN-2 (cortical neuronal cell). Over 14 days, cells were exposed to EMF for 1, 3, or 6 hours per day (hrs/d). The results pointed to a cell type-specific reaction to EMF exposure. In addition, the cellular responses were dependent on duration of EMF exposure. In the present study, cell proliferation was the trait most sensitive to EMF. EMF treatment promoted growth of hFOB 1.19 and HCN-2 compared with control cells at 7 and 14 days of incubation. When the exposure time was 3 hrs/d, EMF enhanced the proliferation of RPMI 7666 but inhibited that of T/G HA- VSMC. On the other hand, the effects of EMF on cell cycle distribution, cell differentiation, and actin distribution were unclear. Furthermore, we hardly found any correlation between EMF exposure and gap junctional intercellular communication in hFOB 1.19. This study revealed that EMF might serve as a potential tool for manipulating cell proliferation.
Signal Transduction ; Microfilaments/radiation effects ; Humans ; Gap Junctions/metabolism/radiation effects ; *Electromagnetic Fields ; Cell Proliferation/radiation effects ; Cell Physiology/*radiation effects ; Cell Line ; Cell Differentiation/radiation effects ; Cell Cycle/radiation effects

No abstract available.
Angiomyoma

SUMMARY: A 21-year-old healthy Korean man worked on a building construction site every day for almost 2 months and exercised every day for 1 or 2 hours after working hard. He felt dizziness, nausea, and experienced vomiting and body aches immediately after exercise and immediately took cold medicines including acetaminophen, cimetidine, bepotastine, and Codenal? complex for the common cold symptoms for 2 days because he was scheduled to participate in navy training at that time. He complained of severe trapezius pain and aches in his left calf 3 days after joining the Navy training. Testing revealed creatine phosphokinase (CPK) 6260 U/L, myogloblin 176 mcg/L in the urine, liver enzymes increased, and oliguria, suggesting rhabdomyolysis. He recovered with intravenous fluids without any complications.
Acetaminophen ; Cimetidine ; Common Cold ; Creatine Kinase ; Dizziness ; Humans ; Liver ; Nausea ; Oliguria ; Rhabdomyolysis* ; Superficial Back Muscles ; Vomiting ; Young Adult

BACKGROUND: Dermatofibrosarcoma protuberans is a mesenchymal tumor of the skin of intermediate-grade which is a rare condition. The slow growing and aggressive invasion on local tissues are characteristic features of dermatofibrosarcoma protuberans. The treatment for dermatofibrosarcoma protuberans is mainly a surgical excision such as a wide excision and Mohs micrographic surgery. OBJECTIVE: The aim of this study was to compare the result of wide excision and Mohs micrographic surgery for the treatment of dermatofibrosarcoma protuberans at a single institution in Korea. METHODS: A retrospective review was done for 24 patients diagnosed with dermatofibrosarcoma protuberans and treated surgically from 1999 to 2010 at Chonbuk National University Hospital. Patient demographics, tumor features, surgical features, and recurrence during the follow-up period were evaluated. RESULTS: 13 patients were treated with wide excision, and 11 with Mohs micrographic surgery. There was no metastasis for all the cases. Mean operation time for the wide excision group was 83 minutes whereas 182 minutes for the Mohs micrographic surgery group, and it was a statistically significant difference. However, no significant difference was observed in post-operative defect size, advanced surgical repair and local recurrence in our study. CONCLUSION: We suggest that wide excision and Mohs micrographic surgery are both successful modalities for the surgical treatment of dermatofibrosarcoma protuberans. Hence, individualized patient and tumor characteristics should be concerned when determining the surgical options for dermatofibrosarcoma protuberans.
Demography ; Dermatofibrosarcoma ; Follow-Up Studies ; Humans ; Mohs Surgery ; Neoplasm Metastasis ; Recurrence ; Retrospective Studies ; Skin

BACKGROUND: A recent advancement in microsurgery, the free flap is widely used in the reconstruction of the lower leg and foot. The simple and effective methods of local flaps, including transposition and advancement flaps, have been considered for patients with chronic debilitation who are unable to endure long surgical procedures or general anesthesia. However, the location and size of the wound may restrict the clinical application of a local flap. Under these circumstances, a sural flap can be an excellent alternative, rendering satisfying clinical outcomes in chronically debilitated patients. METHODS: Between 2008 and 2012, 39 patients underwent soft tissue defect treatment by sural artery flap as a final method. All of the patients had at least one chronic disease or more (diabetes, hypertension, vascular disease, etc.). Also, all of the patients had a history of chronic lower extremity ulceration, which revealed no response to several months of conservative treatment. RESULTS: The results of the 39 cases had a success rate of 100% with 39 complete recoveries. Nine cases suffered complications: partial necrosis (n=4), wound dehiscence without necrosis (n=3), hematoma (n=1), and infection (n=1). CONCLUSIONS: The sural artery flap is not only useful for the lower leg but also for the heel, and other various parts. Furthermore, it is a relatively simple surgical technique for reconstructing the defect area for patients with various chronic conditions with a high surgical risk or contraindications to surgery.
Anesthesia, General ; Arteries ; Chronic Disease ; Foot ; Free Tissue Flaps ; Heel ; Hematoma ; Humans ; Hypertension ; Leg ; Lower Extremity ; Microsurgery ; Necrosis ; Organic Chemicals ; Soft Tissue Injuries ; Surgical Flaps ; Ulcer ; Vascular Diseases

STUDY DESIGN: In vitro experimental study. OBJECTIVES: To examine the cellular proliferation, synthetic activity and phenotypical expression of intervertebral disc (IVD) cells seeded on types I and II atelocollagen scaffolds, with the stimulation of TGF-beta1 and BMP-2. SUMMARY OF LITERATURE REVIEW: Recently, tissue engineering is regarded as a new experimental technique for the biological treatment of degenerative IVD diseases, and has been highlighted as a promising technique for the regeneration of tissues and organs in the human body. Research on cell transplantation in artificial scaffolds has provided that the conditions for tissue engineering have to be equilibrated, including the cell viability and proliferation, maintenance of characteristic phenotype, suitable scaffolds in organisms and biologically stimulated growth factor. MATERIAL AND METHOD: Lumbar IVD cells were harvested from 10 New Zealand white rabbits, with the nucleus pulposus cells isolated by sequential enzymatic digestion. Each of 1% types I and II atelocollagen dispersions were poured into a 96-well plate (diameter 5 mm), frozen at -70 degrees C, and then lyophilized at -50 degrees C. Fabricated porous collagen matrices were made using the cross-linking method. Cell suspensions were imbibed by surface tension into a scaffold consisting of atelocollagen. The cell cultured scaffolds were then treated with TGF-beta1 (10 ng/ml) or BMP-2 (100 ng/ml) or both. After 1 and 2 week culture periods, the DNA synthesis was measured by [3H] thymidine incorporation, and newly synthesized proteoglycan by incorporation of [35S] sulphate. Reverse transcription-polymerase chain reactions for the mRNA expressions of type I and II collagen, aggrecan and osteocalcin were performed. The inner morphology of the scaffolds was determined by scanning electron microscopy (SEM). RESULTS: The IVD cultures in collagen type II with TGF-beta1 demonstrated an increase in proteoglycan synthesis and up regulation of aggrecan and types I and II collagen mRNA expressions compared to the control. IVD cultures in the type I atelocollagen scaffold with growth factors exhibited an increase in DNA synthesis and up regulation of the type II atelocollagen mRNA expression. With all combinations of growth factor, the IVD cultures in types I and II atelocollagen scaffolds showed no up regulation of the osteocalcin mRNA expression. Furthermore, there was no synergistic effect of TGF-beta1 and BMP-2 in the matrix synthesis or for the mRNA expression of the matrix components. CONCLUSIONS: Nucleus pulposus cells from rabbit were viable in atelocollagen types I and II atelocollagen scaffolds. The type I atelocollagen scaffold was suitable for cell proliferation, but the type II atelocollagen scaffold was more suitable for extracellular matrix synthesis. The IVD cells in both scaffolds were biologically responsive to growth factors. Taken together, nucleus pulposus cells in atelocollagen scaffolds, with anabolic growth factors, provide a mechanism for tissue engineering of IVD cells.
Aggrecans ; Cell Proliferation ; Cell Survival ; Cell Transplantation ; Collagen ; Collagen Type II ; Digestion ; DNA ; Extracellular Matrix ; Human Body ; Intercellular Signaling Peptides and Proteins* ; Intervertebral Disc* ; Microscopy, Electron, Scanning ; Osteocalcin ; Phenotype ; Proteoglycans ; Rabbits ; Regeneration ; RNA, Messenger ; Surface Tension ; Suspensions ; Thymidine ; Tissue Engineering ; Transforming Growth Factor beta1 ; Transplants ; Up-Regulation

STUDY DESIGN: In vitro experimental study. OBJECTIVES: To examine the cellular proliferation, synthetic activity and phenotypical expression of intervertebral disc (IVD) cells seeded on types I and II atelocollagen scaffolds, with the stimulation of TGF-beta1 and BMP-2. SUMMARY OF LITERATURE REVIEW: Recently, tissue engineering is regarded as a new experimental technique for the biological treatment of degenerative IVD diseases, and has been highlighted as a promising technique for the regeneration of tissues and organs in the human body. Research on cell transplantation in artificial scaffolds has provided that the conditions for tissue engineering have to be equilibrated, including the cell viability and proliferation, maintenance of characteristic phenotype, suitable scaffolds in organisms and biologically stimulated growth factor. MATERIAL AND METHOD: Lumbar IVD cells were harvested from 10 New Zealand white rabbits, with the nucleus pulposus cells isolated by sequential enzymatic digestion. Each of 1% types I and II atelocollagen dispersions were poured into a 96-well plate (diameter 5 mm), frozen at -70 degrees C, and then lyophilized at -50 degrees C. Fabricated porous collagen matrices were made using the cross-linking method. Cell suspensions were imbibed by surface tension into a scaffold consisting of atelocollagen. The cell cultured scaffolds were then treated with TGF-beta1 (10 ng/ml) or BMP-2 (100 ng/ml) or both. After 1 and 2 week culture periods, the DNA synthesis was measured by [3H] thymidine incorporation, and newly synthesized proteoglycan by incorporation of [35S] sulphate. Reverse transcription-polymerase chain reactions for the mRNA expressions of type I and II collagen, aggrecan and osteocalcin were performed. The inner morphology of the scaffolds was determined by scanning electron microscopy (SEM). RESULTS: The IVD cultures in collagen type II with TGF-beta1 demonstrated an increase in proteoglycan synthesis and up regulation of aggrecan and types I and II collagen mRNA expressions compared to the control. IVD cultures in the type I atelocollagen scaffold with growth factors exhibited an increase in DNA synthesis and up regulation of the type II atelocollagen mRNA expression. With all combinations of growth factor, the IVD cultures in types I and II atelocollagen scaffolds showed no up regulation of the osteocalcin mRNA expression. Furthermore, there was no synergistic effect of TGF-beta1 and BMP-2 in the matrix synthesis or for the mRNA expression of the matrix components. CONCLUSIONS: Nucleus pulposus cells from rabbit were viable in atelocollagen types I and II atelocollagen scaffolds. The type I atelocollagen scaffold was suitable for cell proliferation, but the type II atelocollagen scaffold was more suitable for extracellular matrix synthesis. The IVD cells in both scaffolds were biologically responsive to growth factors. Taken together, nucleus pulposus cells in atelocollagen scaffolds, with anabolic growth factors, provide a mechanism for tissue engineering of IVD cells.
Aggrecans ; Cell Proliferation ; Cell Survival ; Cell Transplantation ; Collagen ; Collagen Type II ; Digestion ; DNA ; Extracellular Matrix ; Human Body ; Intercellular Signaling Peptides and Proteins* ; Intervertebral Disc* ; Microscopy, Electron, Scanning ; Osteocalcin ; Phenotype ; Proteoglycans ; Rabbits ; Regeneration ; RNA, Messenger ; Surface Tension ; Suspensions ; Thymidine ; Tissue Engineering ; Transforming Growth Factor beta1 ; Transplants ; Up-Regulation

BACKGROUND: Telomere shortening is thought to be involved in the pathophysiology of myeloid malignancies, but telomere lengths (TL) during interphase and metaphase in hematopoietic malignancies have not been analyzed. We aimed to assess the TLs of interphase and metaphase cells of MDS and telomerase activity (TA) and to find out prognostic significances of TL and TA. METHODS: The prognostic significance of TA by quantitative PCR and TL by quantitative fluorescence in situ hybridization (QFISH) of interphase nuclei and metaphase chromosome arms of bone marrow cells from patients with MDS were evaluated. RESULTS: MDS patients had shorter interphase TL than normal healthy donors (P<0.001). Average interphase and metaphase TL were inversely correlated (P=0.013, p arm; P=0.029, q arm), but there was no statistically significant correlation between TA and TL (P=0.258). The progression free survival was significantly shorter in patients with high TA, but the overall survival was not different according to average TA or interphase TL groups. Multivariable Cox analysis showed that old age, higher International Prognostic Scoring System (IPSS) subtypes, transformation to AML, no history of hematopoietic stem cell transplantation and short average interphase TL (<433 TL) as independent prognostic factors for poorer survival (P=0.003, 0.001, 0.005, 0.005, and 0.013, respectively). CONCLUSIONS: The lack of correlation between age and TL, TA, and TL, and the inverse relationship between TL and TA in MDS patients reflect the dysregulation of telomere status and proliferation. As a prognostic marker for leukemia progression, TA may be considered, and since interphase TL has the advantage of automated measurement by QFISH, it may be used as a prognostic marker for survival in MDS.
Arm ; Bone Marrow Cells ; Disease-Free Survival ; Fluorescence ; Hematologic Neoplasms ; Hematopoietic Stem Cell Transplantation ; Humans ; In Situ Hybridization ; Interphase ; Leukemia ; Metaphase ; Myelodysplastic Syndromes* ; Polymerase Chain Reaction ; Prognosis ; Telomerase* ; Telomere Shortening ; Telomere* ; Tissue Donors