• The Korean Journal of Physiology and Pharmacology
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• v.16 no.3
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• pp.167-174
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• 2012

Natural killer (NK) cells provide one of the initial barriers of cellular host defense against pathogens, in particular intracellular pathogens. Because bone marrow-derived hematopoietic stem cells (HSCs), lymphoid protenitors, can give rise to NK cells, NK ontogeny has been considered to be exclusively lymphoid. Here, we show that porcine c-$kit^+$ bone marrow cells (c-$kit^+$ BM cells) develop into NK cells in vitro in the presence of various cytokines [interleukin (IL)-2, IL-7, IL-15, IL-21, stem cell factor (SCF), and fms-like tyrosine kinase-3 ligand (FLT3L)]. Adding hydrocortisone (HDC) and stromal cells greatly increases the frequency of c-$kit^+$ BM cells that give rise to $CD2^+CD8^+$ NK cells. Also, intracellular levels of perforin, granzyme B, and NKG2D were determined by RT-PCR and western blotting analysis. It was found that of perforin, granzyme B, and NKG2D levels significantly were increased in cytokine-stimulated c-$kit^+$ BM cells than those of controls. And, we compared the ability of the cytotoxicity of $CD2^+CD8^+$ NK cells differentiated by cytokines from c-$kit^+$ BM cells against K562 target cells for 28 days. Cytokines-induced NK cells as effector cells were incubated with K562 cells as target in a ratio of 100 : 1 for 4 h once a week. In results, $CD2^+CD8^+$ NK cells induced by cytokines and stromal cells showed a significantly increased cytotoxicity 21 days later. Whereas, our results indicated that c-$kit^+$ BM cells not pretreated with cytokines have lower levels of cytotoxicity. Taken together, this study suggests that cytokines-induced NK cells from porcine c-$kit^+$ BM cells may be used as adoptive transfer therapy if the known obstacles to xenografting (e.g. immune and non-immune problems) were overcome in the future.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.1
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• pp.114-122
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• 1995

Defining the mechanisms of B cell diversification which establish the immune repertoire is fundamental to understand how the immune response is regulated. In this report, B cell differentiation and diversification focused on the regulation of immunoglobulin $V_H$ gene expression during ontogeny were analyzed by in situ hybridization technique. Fetal liver B cells in .different gestational days from 16d to 20d showed the predominant expression of $V_H7183$ and $V_HQ52$ without transition of repertoire during the observed gestation days. The two subsets of fetal liver B cells separated according to different differentiation stages based on the presence of tell surface immunoglobulin also did not indicate apparent difference in expressed $V_H$ gene family profiles. B cells in fetal spleen as an another hematopoietic lymphoid tissue in fetus also expressed similar $V_H$ gene repertoire to that in fetal liver B cells. This distinct pattern of $V_H$ gene expression in fetal B cells from that of adult B cells were not changed even after four weeks contact with adult bone marrow microenvironment supplied by the established adult bone marrow stromal cell layers. Thus, the restricted $V_H$ gene repertoire of B cells in fetus which is distinct from that in adult appears to be associated more with the genetic potential of fetal B cell progenitors and less with environmental influences or differentiation stages or compartmentalization.

• Journal of the korean academy of Pediatric Dentistry
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• v.31 no.1
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• pp.79-84
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• 2004

Pilomatricoma is a benign soft tissue tumor of hair follicle origin. They occur most commonly in the head and neck region and are usually found in girls during the first two decades of life. Although malignant transformation has been described, it is exceedingly rare. The clinical presentation is typically that of an asymptomatic, superfical, solitary, firm mass that is often accompanied by a reddish-blue discoloration of the overlying skin. Histopathologically pilomatricoma are seen as epithelial islands embedded in a cellular stroma. The epithelial component consist of two main cell types : basaloid cells and ghost cells. Intracellular and stromal calcification is reported in many cases. Diagnosis is usually suspected based on palpation of a superficial, rock-hard mass and confirmed by histopathologic examination. Surgical excision is both curative and the treatment of choice. Recurrence is rare. In this case, the radiographs showed a dense calcification, measuring about 1cm diameter. So simple excision was peformed intraorally, and resultantly the lesion was proved to be a pilomatricoma by histological examination.

• Korean Journal of Veterinary Research
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• v.36 no.1
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• pp.93-99
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• 1996

This study was designed to investigate the effect of estrogen(Est) on the progestcrone(Prog) target cells by autoradiography. The spayed 16 mice(ICR, approximately 18~25g) were randomly alloted into 3 groups. $^3H$-Prog-treated group were injected with $40{\mu}Ci$ of $^3H$-Prog/mouse/day for 1 day, Est + $^3H$-Prog-treated group with $20{\mu}Ci$ of $17{\beta}$-Est/mouse/day for 3 days and then with $40{\mu}Ci$ of $^3H$-Prog/mouse at 4th day, and Est+$^3H$-thymidine(TdR)-treated group with $20{\mu}g$ of $17{\beta}$-Est/mouse/day for 3 days and then $80{\mu}Ci$ of $^3H$-TdR/mouse at 4th days. 1. Mice uteri of both Est+$^3H$-Prog-treated group and Est+$^3H$-TdR-treated group were hypemophied in gross finding and the endometrium and myometrium were thickened in microscopic findings. These findings were confirmed that Est enlarged the uteri of mice. 2. Cryo-preparations of mice organs were processed for autoradiography using Kodak NTB-2 emulsion following Kodak D-19 developer and hematoxylin counterstain. In each group, the number values of silver grain distribution appeared to be higher in the $^3H$-Prog-treated group than in the Est+$^3H$-Prog-treated group. It was considered that Est and Prog inhibit each other in action. 3. In both $^3H$-Prog-treated group and Est+$^3H$-Prog-treated group, the uteri have highest distribution rates of silver grains than in other organs, and the cerebral neurons, hepatocytes, bronchiolar epithelial cells and splenic reticular cells also contained some silver grains. 4. The orders of the cell types with more number of silver grains in the uteri were stromal cells, glandular epithelial cells, luminal surface cells and muscular cells and also were as above orders in distribution of proliferating cell type by $^3H$-TdR.

• Journal of Veterinary Clinics
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• v.30 no.3
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• pp.214-217
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• 2013

A 3-year-old intact female hedgehog (Atelerix albiventris) was presented for evaluation of mandibular swelling. Fine needle aspiration of swollen mandibular was performed and smears were stained with Romanowsky type stain for cytological evaluation. Smears were highly cellular with predominance of variably shaped keratinized or non keratinized squamous cells with low numbers of spindloid to abnormally elongated cells. Cytologic impression was squamous cell carcinoma. The mass was surgically removed for histological examination. Microscopically tortuous and anastomosing delicate to broad pegs and nests of neoplastic squamous epithelial cells were supported by a moderate collagenous and spindloid fibroblast stroma. Tumor cells had moderate anisocytosis and mild anisokaryosis and range from moderately to well keratinized, with areas of intratumoral acantholysis accompanied by mixed stromal lymphoplasmacytic, neutrophilic inflammation. The mitotic index is 2-3 per high-power field. Tumor cells were expanding the subcutis subjacent to the layer of skeletal muscle and incorporating the osseous tissue fragments. The final diagnosis was squamous cell carcinoma. The patient survived three months after surgery without any further medical treatments.

• Radiation Oncology Journal
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• v.24 no.3
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• pp.164-170
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• 2006

[ $\underline{Purpose}$ ]: This study identified the result of postoperative radiation therapy and the prognostic factors to affect survival rates in cancer patients. $\underline{Materials\;and\;Methods}$: One hundred and thirty three patients with cervical cancer who were treated with postoperative radiation therapy following surgery at our institution between June 1985 and November 2002 were retrospectively analyzed. One hundred and thirteen patients had stage IB disease, and 20 patients had stage IIA disease. Histological examination revealed 118 squamous cell carcinoma patients and 15 adenocarcinoma patients. Sixty seven patients were noted to have stromal invasion greater than 10 mm, and 45 patients were noted to have stromal Invasion 10 mm or less. Positive lymphovascular invasion was found in 24 patients, and positive pelvic lymph nodes were noted in 39 patients. Positive vaginal resection margin was documented in 8 patients. All of the patients were treated with external beam radiation therapy to encompass whole pelvis and primary surgical tumor bed. Intracavitary radiation therapy was added to 19 patients who had positive or close surgical margins. $\underline{Results}$: Actuarial overall and disease-free survival rates for entire group of the patients were 88% and 84% at 5 years, respectively. Five-year disease-free survival rates for patients with stromal invasion greater than 10 mm and 10 mm or less were 76% and 97%, respectively (p<0.05). Also there was a significantly lower survival in patients with positive pelvic lymph nodes compared with patients with negative pelvic lymph nodes (p<0.05). However, lymphovascular invasion, positive vaginal resection margins were not statistically significant prognostic factors. Addition of neoadjuvant chemotherapy or type of surgery did not affect disease-free survival. $\underline{Conclusion}$: Postoperative radiation therapy appears to achieve satisfactory local control with limited morbidity in cervical cancer patients with high pathologic risk factors. Distant metastasis was a dominant failure pattern to affect survival in cervical cancer patients after radical surgery and radiation and more effective systemic treatment should be investigated in these high-risk patients.

• Radiation Oncology Journal
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• v.22 no.4
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• pp.265-270
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• 2004

Purpose: To evaluate the effectiveness of postoperative radiation therapy in cervical cancer patients and define the prognostic factors to affect survival rates. Materials and Methods: Eighty one patients with cervical cancer who were treated with postoperative radiation therapy following surgery at our institution between May 1992 and April 2000 were retrospectivelv analyzed. Forty two patients had stage IB disease, 17 had stage IIA disease, and remaining 22 had stage IIB disease, respectively. Histological examination revealed 76 squamous cell carcinoma and 5 adenocarclnoma. Sixty one patients were noted to have stromal invasion greater than 8 mm and 20 patients were noted to have stromal invasion 7 mm or less. Sixteen patients had parametrial invasion and 65 patients did not. Positive vaginal resection margin was documented in only eight patients and positive lymphovascular invasion was in twelve patients. All of the patients were treated with external beam radiation therapy alone. Majority of the patients were treated with 4 field brick technique to encompass whole pelvis. Total of 5,500 cGy was delivered to the primary surgical tumor bed. Minimum follow up period was four years. Results: Actuarial disease free survival rates for entire group of the patients were 95% and 89% at 2 and 5 years, respectively Five year disease free survival rates for patients with stage IB, IIA, and IIB disease were 97%, 87% and 70%, respectivelv. Local recurrences were documented in 5 patients. Cumulative local failure rate at 3 years was 6% Five year disease free survival rates for patients with stromal invasion greater than 8 mm and 7 mm or less were 88% and 92%, respectively (p>0.05). Five year disease free survival rate for patients with parametrial invasion was significantly lower than those with no invasion (72% vs 92%, p<0.05). Also there was significantly lower survival in patients with positive vaginal resection margin, compared with patients with negative resection margin (64% vs 94%, p<0.05). However, lymphovascular invasion was not a statistically significant prognostic factor Parametrial invasion and positive surgical resection margins were noted to be significant prognostic factors. Conclusions: Postoperative radiation therapy appears to be beneficial in controlling local disease in cervical cancer patients with high pathologic risk factors. Parametrial invasion and positive resection margins were noted to be significant prognostic factors to affect survival and more effective treatment should be investigated in these patients.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.4
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• pp.507-515
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• 2014

Benign prostatic hyperplasia (BPH), which is commonly found in aging men, is characterized by hyperplasia of prostatic stromal and epithelial cells beginning in the periurethral zone of the prostate. The prevalence of BPH increases in an age-dependent manner. Here, we investigated the protective effects of unripe Rubus occidentalis extracts (UROE) on BPH development using a prostate cancer cell line and testosterone-induced BPH rat model. Experiments using an established hormone-dependent prostate cancer cell line (LNCaP) showed that UROE treatment significantly decreased expression of androgen-related genes, including androgen receptor (AR), prostate specific antigen (PSA), and 5-alpha reductase 2, but not 5-alpha reductase 1, which was also observed in flutamide-treated cells. Further, AR and PSA gene expression was reduced by UROE treatment under androgen-stimulated conditions using dihydrotestosterone (DHT). BPH animals displayed elevated prostate weights. However, UROE as well as finasteride treatment significantly reduced prostate weights and DHT levels compared to testosterone-induced BPH animals. Histopathological analysis also showed that UROE treatment suppressed testosterone-induced prostatic hyperplasia. Taken together, the results suggest that UROE may effectively inhibit the development of BPH and thus may be a useful agent in BPH treatment.

• Development and Reproduction
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• v.14 no.4
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• pp.233-241
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• 2010

Human adipose stem cells are an abundant, readily available population of multipotent progenitor cells that reside in adipose tissue and these cells have characteristics very similar to bone marrow mesenchymal stromal cells (BMMSCs). However, liposuction procedure, donor age, body mass index, and harvesting sites might generate differences in the initial cell population and the preparations are a heterogeneous mixture of precursors with different subsets. Therefore, in this study, we investigated the characteristics of human thigh adipose stem cells and the differentiation potential into mesodermal and endodermal lineage. Thigh adipose stem cells maintained fibroblast-like morphology similar to BM-MSCs and they underwent average 56.5 doublings and produced $5{\times}10^{22}$ cells. These cells expressed SCF, Oct4, nanog, vimentin, CK18, FGF5, NCAM, Pax6, BMP4, HNF4a, nestin, GATA4, HLA-ABC, and HLA-DR genes at p3 and they also expressed Oct4, Thy-1, FSP, vWF, vimentin, desmin, CK18, CD54, CD4, CD106, CD31, a-SMA, HLA-ABC proteins. Moreover, they could differentiate into mesodermal lineage cells such as adipocyte, osteoblast and chondrocyte. In addition, they also differentiated into insulin secreting cells in our culture condition. In conclusion, human thigh adipose stem cells retain proliferative potential and expression patterns similar to BM-MSCs and they also differentiate into various cell types. Thus, human thigh adipose stem cells might be useful alternative cell source for clinical application.

• Journal of Periodontal and Implant Science
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• v.39 no.sup2
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• pp.279-286
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• 2009

Purpose: Gene therapy (ex vivo) has recently been used as a means of delivering bone morphogenetic proteins (BMPs) to sites of tissue regeneration. In the present study, we investigated the effect of co-transduction of adenoviruses expressing BMP-2 and BMP-7 on osteogenesisof C2C12 cells in vitro. Methods: A replication-defective human adenovirus 5 (Ad5) containing a cDNA for BMPs in the E1 region of the virus (Ad5BMP-2 and Ad5BMP-7) was constructed by in vivo homologous recombination. Functional activity of Ad5BMP-2 and Ad5BMP-7 were evaluated in mouse stromal cells (W20-17cells). C2C12 cells are transduced with various MOI (multiplicity of infection) of Ad5BMP-2 and Ad5BMP-7 to assess most effective and stable titer. Based on this result, C2C12 cells were transduced with Ad5BMP-2 and Ad5BMP-7 alone or by combination. BMPs expression, alkaline phosphatase (ALPase) activity, cell proliferation, and mineralization were assessed. Results: Ad5BMP-2 and Ad5BMP-7 are successfully transduced to W20-17 cells, and secreted BMPs stimulated cell differentiation. Also, C2C12 cells transduced with Ad5BMPs showed expression of BMPs and increased ALPaseactivity. In all groups, cell proliferation was observed over times. At 7days, cells co-transduced with Ad5BMP-2 and Ad5BMP-7 showed lower proliferation than the others. C2C12 cells co-transduced with Ad5BMP-2 and Ad5BMP-7 had greater ALPaseactivity than that would be predicted if effect of individual Ad5BMPs were additive. Little mineralized nodule formation was detected in cells transduced with individual Ad5BMPs. In contrast, Ad5BMP-2 and Ad5BMP-7 combination stimulated mineralization after culturing for 10 days in mineralizing medium. Conclusions: Present study demonstrated that adenoviruses expressing BMPs gene successfully produced BMPs protein and these BMPs stimulated cells to be differentiated into osteoblastic cells. In addition, the osteogenic activity of Ad5BMPs can be synergistically increased by co-transduction of cells with Ad5BMP-2 and Ad5BMP-7.