• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.4
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• pp.299-305
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• 2010

Purpose: Adipose tissue is located beneath the skin, around internal organs, and in the bone marrow in humans. Its main role is to store energy in the form of fat, although it also cushions and insulates the body. Adipose tissue also has the ability to dynamically expand and shrink throughout the life of an adult. Recently, it has been shown that adipose tissue contains a population of adult multipotent mesenchymal stem cells and endothelial progenitor cells that, in cell culture conditions, have extensive proliferative capacity and are able to differentiate into several lineages, including, osteogenic, chondrogenic, endothelial cells, and myogenic lineages. Materials and Methods: This study focused on endothelial cell culture from the adipose tissue. Adipose tissues were harvested from buccal fat pad during bilateral sagittal split ramus osteotomy for surgical correction of mandibular prognathism. The tissues were treated with 0.075% type I collagenase. The samples were neutralized with DMEM/and centrifuged for 10 min at 2,400 rpm. The pellet was treated with 3 volume of RBC lysis buffer and filtered through a 100 ${\mu}m$ nylon cell strainer. The filtered cells were centrifuged for 10 min at 2,400 rpm. The cells were further cultured in the endothelial cell culture medium (EGM-2, Cambrex, Walkersville, Md., USA) supplemented with 10% fetal bovine serum, human EGF, human VEGF, human insulin-like growth factor-1, human FGF-$\beta$, heparin, ascorbic acid and hydrocortisone at a density of $1{\times}10^5$ cells/well in a 24-well plate. Low positivity of endothelial cell markers, such as CD31 and CD146, was observed during early passage of cells. Results: Increase of CD146 positivity was observed in passage 5 to 7 adipose tissue-derived cells. However, CD44, representative mesenchymal stem cell marker, was also strongly expressed. CD146 sorted adipose tissue-derived cells was cultured using immuno-magnetic beads. Magnetic labeling with 100 ${\mu}l$ microbeads per 108 cells was performed for 30 minutes at $4^{\circ}C$ a using CD146 direct cell isolation kit. Magnetic separation was carried out and a separator under a biological hood. Aliquous of CD146+ sorted cells were evaluated for purity by flow cytometry. Sorted cells were 96.04% positivity for CD146. And then tube formation was examined. These CD146 sorted adipose tissue-derived cells formed tube-like structures on Matrigel. Conclusion: These results suggest that adipose tissue-derived cells are endothelial cells. With the fabrication of the vascularized scaffold construct, novel approaches could be developed to enhance the engineered scaffold by the addition of adipose tissue-derived endothelial cells and periosteal-derived osteoblastic cells to promote bone growth.

• Tuberculosis and Respiratory Diseases
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• v.50 no.6
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• pp.693-703
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• 2001

Background : Smoking is the most important and consistent determinant of the development and progression of COPD(Ed Note : Define COPD). The fact that cigarette smokers develop a different type of COPD, chronic bronchitis and emphysema, with different clinical and pathological aspects, suggests that the development of COPD has a relationship with other smoking-associated factors beyond just a simple smoking history. The aim of this was to analyze the smoking habits and history of patients with COPD and to evaluate the development of different types of COPD according to patient's smoking habits. Method : To evaluate the differences in the smoking patterns of patients with chronic bronchitis and emphysema, a pulmonary function test was conducted, and the smoking history and patterns was obtained through a smoking history questionnaire by a direct personal interview from 333 male cigarette smokers diagnosed with COPD, in the Yeungnam university medical center(190 patients diagnosed with chronic bronchitis, 143 patients diagnosed with emphysema). Result : The patients with emphysema smoked earlier and had a higher smoking history(ie, more packyears, more total amounts of smoked cigarette, and more deep inhalation and longer duration of plain cigarette exposure) than those with chronic bronchitis. The depth of inhalation was also significantly higher in the emphysema patients after taking into account age, cumulative cigarette consumption and the type of cigarette smoked. Conclusion : Emphysema was more associated with the increasing degree of inhalation as assessed by the depth of inhalation. A high alveolar smoke exposure may be a significant risk factor for the development of emphysema.

• The Journal of the Korean bone and joint tumor society
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• v.9 no.2
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• pp.190-199
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• 2003

Purpose: The aim of this study was to find out a clinically appliable method to insert a biodegradable solid material containing holmium-166-chitosan complex into the surgical field, and to evaluate the histological changes in the normal tissues after ${\beta}$ -ray irradiation from holmium-166 according to the dose, period and type of tissues. Materials and Methods: 3.0 mCi, 50 ${\mu}l$ of the liquid state $^{166}$Ho-chitosan complex was attached to the absorbable gelatin sponge. The radiation activity measured by dose caliberator was 1.5 mCi. These $^{166}$Ho-chitosan complex containing absorbable gelatin sponges were inserted into the thigh muscles and over the femur bones of the Wistar rats. The cases were evaluated at 2 weeks after insertion, and 4, 6 weeks with respect to the histological changes of the soft tissues and bone, the depth of the tissue necrosis, and the changes of the $^{166}$Ho-chitosan complex containing absorbable gelatin sponges. Results: At 2 weeks, the muscles showed coagulation necrosis, degenerating myocytes, regenerating myocytes, intermuscular edema, and inflammatory cells. The necrosis depth was 3.3 mm. In the bones, there was no osteocyte in the lacuna of cortex (empty lacuna), marrow fibrosis, inflammation. The necrosis depth was 2.9 mm. At 4 weeks, in the muscle, calcification and increased fibrosis with necrosis depth by 3.3 mm were the additional findings. In the bone, marrow fibrosis with necrosis depth by 3.3 mm were detected. At 6 weeks, soft tissue shrinkage, increased fibrosis and granulation tissue formation, and nearly resolving inflammatory reaction were the findings. Conclusion: The local application of the $^{166}$Ho-chitosan complex attached to biodegradable gelatin material with surgery in the laboratory animals resulted in no mortality and morbidity, and satisfactory tissue necrosis. Holmium-166 can be applied to the treatment of the malignant tumor patients.

• Pediatric Infection and Vaccine
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• v.15 no.2
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• pp.180-187
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• 2008

Purpose : We wanted to determine the characteristics of patients with Kawasaki disease (KD) who were unresponsive to intravenous immunoglobulin (IVIG). Methods : The patients with KD were divided into two groups: the IVIG responsive group (25 cases) and the IVIG unresponsive group (14 cases). We analyzed various parameters before and after the administration of IVIG, including the complete blood cell count with the differential count (%), the erythrocyte segmentation rate (ESR), the C-reactive protein (CRP) level and the protein and lipid profiles. Results : The IVIG unresponsive group had a prolonged duration of fever and a higher incidence of CAL compared to the IVIG responsive group (P<0.001, respectively). Before IVIG infusion, the neutrophil differential, the ESR and the CRP values were higher (P<0.001), and the total protein and albumin values were lower in the IVIG unresponsive group (P=0.01) compared to the IVIG responsive group. After IVIG infusion, there were no significant changes in the WBC count and CRP levels in the IVIG unresponsive group. The reduction of the HDL-cholesterol levels by IVIG was more significant in the unresponsive group (P=0.02). Conclusion : A more severe and prolonged inflammatory response occurred in the IVIG unresponsive group at an early stage, and this finding can be detected by such inflammatory parameters as the neutrophil count and the CRP and HDL-cholesterol levels after IVIG infusion.

• Journal of Life Science
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• v.18 no.11
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• pp.1584-1591
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• 2008

The effect of Chlorella hot water extract (CE) on hyperglycemia in streptozotocin- induced diabetic rats has not been studied. Therefore, hypoglycemic effect of CE in type I streptozotocin- induced diabetic rats was studied. Rats were fed a semisynthetic diet supplemented with either 3% (the STZ+CE3) and 6% (the STZ+CE6) CE or no supplement the Normal and the STZ-Control rats for 4 weeks. The concentrations of fasting and non-fasting blood glucose were higher in the STZ-Control rats than in the Normal rats, but this rise was lowered in the STZ+CE3 and the STZ+CE6 rats. Serum insulin concentrations were decreased with STZ injection, however, the decreased levels were almost restored to the Normal level with CE supplementation. The increased serum fructosamine levels associated with hyperglycemia were decreased with the CE treatment. The morphology of pancreatic islets in the Normal rat was round and maintained a typical arrangement. The STZ-Control pancreatic beta-cells were found to have significant swelling and severely morphological damaged, however, pancreatic tissue damage by STZ in the CE-supplemented diet group was ameliorated. This study shows that Chlorella hot water extract had a hypoglycemic effect on the STZ-diabetic rats via either increased insulin secretion during recovery or the prevention of STZ-induced pancreatic damage.

• Korean journal of food and cookery science
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• v.21 no.5
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• pp.725-732
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• 2005

This study attempted to investigate if the soy isoflavone, genistein, influences bone metabolism in ovariectomized, 4-week-old female Wister rats. All the rats were divided into sham (SH) and ovariectomized (OVX) groups consisting of OVX-17${\beta}$-estradiol($10\;{\mu}g/kg$ b.w.), OVX-1mg or 5 mg or 10mg of genistein/kg b.w. The rates were allowed ad libitum access to foods and water for 8 weeks. The Results showed that body weight had significantly increased in the OVX group compared to the SH group (p<0.05) and was not different among the OVX-GEN and OVX-ES groups and the OVX group. The liver and uterus weights in the OVX groups were slighter than those in SH group (p<0.05). The kidney weight in the OVX groups was decreased compared to in that in the SH group but was not significantly different among all OVX groups. Femoral length in the OVX groups was longer than in the SH group and was not different. Rats in the OVX groups had higher creatinine levels than those in the SH group and hydroxyproline level did not differ significantly among any of the groups. Serum ALP activity and Ca in bone, feces, urine and serum did not change among the groups. Bone mineral density (BMD) decreased in the OVX groups compared to the SH group and was slightly increased by feeding genistein (p>0.05). Breaking force and stiffness did not change by ovariectomy and feeding genistein. Hence, these results suggested that estrogen may affect bone mineralization in growing rats and that genistein be may involved in the prevention of bone loss. However, more studies are needed to identify the proper mechanism of genistein and bone formation.

• Development and Reproduction
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• v.5 no.2
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• pp.175-180
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• 2001

The ectopic expression of gonadotropin releasing hormone(GnRH and luteinizing hormone(LH) in several tissues is a quite intriguing phenomenon. Recently, the presence of GnRH and its receptor has been clearly demonstrated in rodents and human mammary gland. In this context, one can postulate that the presence of local circuit composed of GnRH and LH in the gland. The present study was undertaken to elucidate whether there is a correlation between the LH expression in rat mammary gland and physiological status during the process of mammary differentiation. LH contents in mammary gland from cycling to weaning rats were measured by radioimmunoassay(RIA). In cycling rats, changes of the LH level in both serum and mammary gland showed similar pattern as the highest level in proestrus and the lowest level in diestrus II stage. While the serum LH levels were fluctuated from pregnant through involution stage, a sharp decline of mammary LH contents was observed in the lactating rats. This decrement was recovered in involuting rats to the level of proestrus stage. Reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analyses demonstrated that the transcriptional activities of the mammary LH and GnRH were increased from diestrus I stage to estrus stage, and the increased levels were maintained in pregnant, lactation and involution stages. To test the hypothesis that the alteration in mammary LH expression might be steroid-dependant, ovariectomy(OVX) and steroid supplement model was employed. As expected, supplement of estradiol(E$_2$) after OVX remarkably decreased serum LH level compared to that in serum from vehicle-only treated rats. Likewise, administration of E$_2$ significantly reduced the mammary LH content. The present study demonstrated that (i) the LH expression in mammary gland could be altered by some physiological parameters such as estrous cycle, pregnancy, lactation and involution, and (ii) ovarian steroid especially estrogen seems to be one of major endocrine factors which are responsible for regulation of mammary LH expression.

• Journal of Periodontal and Implant Science
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• v.27 no.4
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• pp.867-882
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• 1997

Safflower(Carthamus tinctorius $Linn\acute{e}$ has been traditionally used for the treatment of blood stasis, and Dipsasi Radix has been used as a drug for fracture in Chinese medicine. The purpose of present study was to examine the biologic effects of safflower extract and Disasi radix extracts on the periodontal. ligament cells and osteoblastic cells and on the wound healing of rat calvarial defect. The ethanolic extract of safflower blossom, safflower seed and Dipsasi Radix(125, 250, and 500 ${\mu}g/ml$) were prepared as test group, and PDGF-BB(lOng/ml) and unsafonifiable fraction of Zea Mays L.(125, 250, and 500 ${\mu}g/ml$) were employed as positive control. The effects of each agents on the growth and survival, ALPase activity, expression of PDGF-BB receptor, chemotactic response of PDL cell and ATCC human osteosarcoma MG63 cells in vitro were examined. The tissue regenerative effect of each extracts was evaluated by histomorphometric measuring of newly formed bone on the 8mm defect in rat calvaria after oral administration of 3 different dosages groups : 0.02, 0.1 and 0.35g/kg, per day. It was also employed the same dosages of unsaponifiable fraction of Zea Mays L. as positive controls. Safflower blossom extract, safflower seed extract, and Dipsasi Radix extract stimulate the cellular activity of MG63 cells in concentration range of $125-500{\mu}g/ml$, and safflower bolssom extract and safflower seed extract stimulate also the cellular activity of periodontal ligament cells in concentration range of $250-500{\mu}g/ml$. In activity of ALPase, $250-500{\mu}g/ml$ of safflower blossom extracts showed significant stimulating effects on MG63 cells, and the same concentration range of safflower seed extracts showed significant effect on periodontal ligament cells. In the recovery on PDGF-BB receptor expression which was depressed by $IL-1{\beta}$, $125-250{\mu}g/ml$ of safflower blossom extracts and $250-500{\mu}g/ml$ of safflower seed extracts showed significant increasing effect on MG63 cells, and $500{\mu}g/ml$ of safflower blossom extract and $250-500{\mu}g/ml$ of safflower seed extracts showed significant effect on periodontal ligament cells. In chemotactic response, among all tested group, safflower seed extracts only were chemotactic to MG63 cells and periodontal ligament cells in concentration range of $125-500{\mu}g/ml$. Also in the view of bone regeneration in rat calvarial defect model, the only group that was orally administrated 0.35g/kg, day of safflower seed extract showed significant new bone formation. These results suggested that safflower extracts might have a potential possibilities as an useful drug for adjunct to treatment for regeneration of periodontal defect.

• Korean Journal of Microbiology
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• v.43 no.3
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• pp.151-158
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• 2007

The possibility of inadvertent introduction of therapeutic gene expressing viral vectors has raised safety concerns about germ-line infection. Particularly, for indications such as prostate cancer and ovarian cancer, the proximity of the point of viral administration to organs of the reproductive system raises concerns regarding inadvertent germ-line transmission of genes carried by the virus vector. To evaluate the safety of in vivo adenovirus mediated gene transfer, we explored the biodistribution, persistance and potential germ-line transmission of p53-expressing adenovirus (Ad-CMV-p53). Both male and female Balb/c mice were injected with $1{\times}10^9$ PFU of Ad-CMV-p53. The PCR analysis showed that there were detectable vector sequences in liver, kidney, spleen, seminal vesicle, epididymis, prostate, ovary, and uterus. The RT-PCR analysis for detecting inserted gene, p53 showed that Ad-CMV-p53 viral RNA were present in spleen, prostate and ovary. Direct injected male and female mice of adenovirus vector into testis and ovary were mated and their of offspring were evaluated for germ-line transmission of the adenoviral vector. The PCR and RT-PCR analysis showed no evidence of germline transmission, although vector sequences were detected in DNA extracted from gonadal tissues. Real-time PCR result confirmed a significant decrease of adenovirus in gonad tissues 1 week after injection. We have also analysed the cell specific localization of viral DNA in gonad tissues by using in-situ PCR. Positive signals were detected in interstitial tissue but not in seminiferous tubule in sperm. In the case of ovary, adenovirus signal were localized to the stromal tissue, but no follicular signals were observed. Together, these data provide strong evidence that the risk of the Inadvertent germ-line transmission of vector sequences following intraperitoneal or direct injection into genito-urinary system of adenovirus is extremely low.

• The Korean Journal of Mycology
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• v.13 no.3
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• pp.169-177
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• 1985

The studies were carried out to obtain the basic data for maximizing the protoplast yields from the mycelia of P. ostreatus and P. sajor-caju. Some factors affecting the regeneration of the protoplast of both species and the productivity of their reversion were also examined. The maximum yields of protoplasts were obtained from four days cultured mycelia of both species on cellophan membrane placed on the surface of PSA or MCM media in a petri dish. The optimal concentration of lytic enzyme Novozym 234 for protoplast releasing was 5 mg per ml of 0.5 M phosphate buffer solution with 0.6 M sucrose or 0.6 M $MgSO_4$ at pH 6.0. The greatest number of protoplasts was released 3 hours after incubation of the mycelia of P. ostreatus and after 4 hours for the P. sajor-caju in the lytic enzyme solution. Among the osmotic stabilizer solutions tested 0.6 M sucrose and 0.6 M KCl showed the best regeneration rates of the protoplasts of both species. When 0.75 % agar solution was over-layed on the regeneration media immediately after inoculation of the protoplast the regeneration rates were greatly enhanced. The ampicillin added to the agar solution prevented bacteria from infection. The reverted isolates produced the sporophores and basidial spores just like their parents without any mutations when they were cultivated in a broad mouth bottle with sawdust substrates.