• Korean Journal of Veterinary Research
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• v.39 no.3
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• pp.628-634
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• 1999

We have studied, by a nonisotopic in situ end-labeling(ISEL) technique, frequency of apoptosis in the external granular layer(EGL) of the cerebellum of immature mice by ${\gamma}$-rays irradiation from $^{60}Co$ or diagnostic ultrasound exposure. The total number of normal cells and cells showing morphological features of apoptosis were counted. The frequency of apoptotic cells was expressed as a percentage of the total number of cells in EGL. The extent of changes following 200 cGy(1090 cGy/min) was studied at 2, 4, 6, 8, 12, or 24 hours after exposure. The maximal frequency was found 6~8 hours after exposure. The immature mice that received 18, 36, 54, 108, 198, 396 cGy of ${\gamma}$-rays or diagnostic ultrasound(7.5MHz, 4.2mW, $I_{SPTA}=7.9mW/cm^2$, $I_{SPTA}=114.3W/cm^2$) for 10 or 30 minutes were examined 6 hours after irradiation. Measurements performed after ${\gamma}$-ray irradiation showed a dose-related increase in apoptotic cells in each of the mice studied. The dose-response curves were analyzed by a linear-quadratic model ; frequency of apoptotic cell in the EGL was y = $(0.1349{\pm}0.01175)D$+$(-0.0001522{\pm}0.0000334)D^2$+0.048($r^2$ = 0.981, D = dose in cGy). In the experiment of ultrasound exposure, the frequency of apoptotic cell was $0.106{\pm}0.130$(10 minutes exposure) and $0.167{\pm}0.220$(30 minutes exposure). We estimated the relative dose of the yield from the experiment with ultrasound by substituting the yield from ultrasound exposure into the curve from the ${\gamma}$-irradiation. The relative dose of ultrasound exposure compared with ${\gamma}$-irradiation were 0.432 cGy(10 minutes exposure) and 0.885 cGy(30 minutes exposure). We have found that there is no evidence to indicate that diagnostic ultrasound involves a significant risk.

• The Journal of Pediatrics of Korean Medicine
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• v.22 no.1
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• pp.69-93
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• 2008

Objectives The purpose of this study is to investigate the effect of KKHS on atopic dermatitis in an in-vivo experiment using an NC/Nga atopic dermatitis mouse, which has histological and clinical similarities to this condition in humans. Methods To investigate the effect of KKHS on atopic dermatitis (AD), we evaluated atopic dermatitis-like skin lesions by clinical skin index and analyzed immunological parameters in peripheral blood mononuclear cells(PBMCs), splenocytes, draining lymph node(DLN) and performed skin histology in ears and dorsal skin of atopic dermatitis of NC/Nga mouse in vivo. Results In vivo, clinical skin severity score was significantly lower in the KKHS group than in the control group. IgE, IL-6, TNF-${\alpha}$, IgM, IgG2a and IgG2b levels in serum decreased remarkably in the KKHS group than in the control group, and the level of IFN-${\gamma}$ production which is secreted from Th1 cell was increased by KKHS. After this experiment we analyzed immunological cells ($CD3^+$, $CD19^+$, $CD4^+$, $CD8^+$, $CD3^+CD69^+$, $CD4^+CD25^+$ and $CD49b^+$) by flow cytometry. It results that the total absolute number of $CD3^+$, $CD19^+$, $CD4^+$ and $CD8^+$ cells were recovered as much as normal state, and the level of $CD3^+CD69^+$ in isolated DLN and PBMCs were significantly decreased, and total absolute number of $Gr-1^+$, $CD11b^+$ and $CD3^+$ in dorsal skin of NC/Nga mouse were decreased by KKHS. We analyzed ear, DLN, and neck-back skin after biopsy and dyeing by hematoxyline/eosin(H&E), toluidine staining (mast cells marker). KKHS were very effective to the histological symptoms which are in dermal and epidermal thickening, hyperkeratosis and inflammatory cell infiltration. Ear thickness was significantly decreased compared with the control group and the size of inflammatory lymphocytes cells (ILC) and plasma cells (PC) in DLN were also decreased. Conclusions KKHS on atopic dermatitis in an in-vivo experiment using an NC/Nga atopic dermatitis mouse was very effectiveness to the atopy dermatitis treatment.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.27 no.4
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• pp.141-157
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• 2014

Objectives : The purpose of this study is to investigate the effects of DOGO phreatic water containing germanium on Atopic Dermatitis in NC/Nga mouse. Methods : We made DOGO phreatic water added germanium. After making atopic dermatitis caused by sensitizing NC/Nga mouse to DNCB(dinitrochlorobenzene), we made mouse swim in tanks each filled with distilled water, tap water, DOGO phreatic water, DOGO phreatic water(added germanium) for 30minutes everyday. 3weeks later, we analyzed skin clinical score, total IgE levels(by ELISA), WBC differential counting(Neutrophils, Monocytes), absolute cell number of $Neutrophil^+Gr-1^+$, CCR3 mRNA expressions(by Real-time PCR), IL-4, IFN-${\gamma}$ production levels(by ELISA), histologic test(by H&E staining, toluidine blue staining). Results : The results of making NC/Nga mouse induced atopic dermatitis swim in tanks filled with DOGO phreatic water(contain germanium) are as follows. 1. Skin clinical scores were decreased significantly in comparison to control group. 2. Total IgG levels were decreased significantly in comparison to control group. 3. WBC differential counting(Neutrophils, Monocytes) were decreased significantly in comparison to control group. 4. Absolute cell number of $Neutrophil^+Gr-1^+$ were decreased significantly in comparison to control group. 5. CCR3 mRNA expressions were decreased significantly in comparison to control group. 6. IL-4, IFN-${\gamma}$ production levels were decreased significantly in comparison to control group. 7. The epithelial tissue thickness, leucocytes infiltration, erythema, edema, excoriation, scaling, mast cells infiltrations in dorsal skin were decreased in comparison to control group. Conclusions : These results indicate that DOGO phreatic water(contain germanium) can be used for helping treat atopic dermatitis.

• The Korea Journal of Herbology
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• v.33 no.4
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• pp.19-26
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• 2018

Objectives : The aim of this study was to investigate the immunopotentiating activity of combine extract that Silkworm and Cinnamomum cassia. Recently, acute epidemic diseases such as cold and viral respiratory diseases have been emerging. So, interested in immunity enhancement has been increasing, and research on natural products to promote immunity activity has been actively conducted. Methods : To confirm the immunopotentiating activity effect, Silkworm (SW), Cinnamomum cassia (CC), and SWCC combined extracts were treated 14 days at 300 mg/kg/day. The changes of glutamic oxalacetic transaminase (GOT), glutamic pyruvate transaminase (GPT) in serum were analyzed after experiment. The changes in the total spleen cell number were measured. Immune cells in spleen were analyzed using fluorescence activated cell sorter (FACS). also, analyzed the expression of cytokines in spleen. Results : Total number of cells in the spleen and FACS analysis of T lymphocytes activated in the spleen showed that the SWCC combined treated group had much higher frequency of active cells than both single groups. The ratio of CD4+CD8+, CD4+CD69+ and CD4+CD25+ T cells in spleen, SWCC is higher than other groups except Nor in CD4+, CD4+CD69+, CD4+CD25+ T cells. The results of this study suggest that SWCC can help immune function via IL-2, IL-10, IL-12, IFN-${\gamma}$ cytokine production, increased T lymphocytes and splenocyte proliferation. Conclusion : Therefore, these results suggested that the SWCC combined extracts administration increase stronger immunity enhancement than when SW and CC adminstration.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.1
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• pp.9-20
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• 1999

The genetic defects in human gametes and embryos can cause adverse effects on overall reproductive events. Biopsy of embryos for preimplantation genetic diagnosis (PGD) offers a new possibility of having children free of the genetic disease. In addition, advanced embryo culture method may enhance the effectiveness of embryo biopsy for the practical application of PGD. This experimental study was undertaken to evaluate the effects of coculture on the development in vitro of biopsied mouse embryos as a preclinical model for PGD of human embryos. Embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BLfemale/CBAmale). Using micromanipulation, 1, 2, 3 or 4 blastomeres of 8-cell stage embryos were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acidic Tyrode's solution (ATS). After biopsy of blastomeres, embryos were cultured in vitro for 110 hours in Ham's F-10 supplemented with 0.4% BSA or cocultured on the monolayer of Vero cells in the same medium. The frequence of blastocyst formation were recorded, and the embryos beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. There was no significant difference in the blastocyst formation between the zona intact control group and the zona drilling (ZD) only, or biopsied groups. The hatching rate of all the treatment groups except 4/8 group was significantly higher than that of control group. In all the treatment groups, there was a significant reduction in the mean cell number of embryos beyond blastocyst stage ($50.2{\pm}14.0$ in control group vs. $41.2{\pm}7.9$ in ZD, $39.3{\pm}8.8$ in 7/8, $29.7{\pm}6.4$ in 6/8, $25.1{\pm}5.7$ in 5/8, and $22.1{\pm}4.3$ in 4/8 groups, p<0.05). When the same treatments were followed by coculture with Vero cells, a similar pattern was seen in the blastocyst formation and the hatching rate. However, in all the treatment groups, there was a significant increase in the mean cell number of embryos beyond blastocyst stage with coculture, compared with the parallel groups without coculture. In the cleavage rate of biopsied blastomeres cultured for 110 hours after IVF, there was no significant difference between coculture and non-coculture groups (87.2% vs. 78.7%). However, the mean cell number of embryos developed from the biopsied blastomeres was significantly higher in coculture group ($11.5{\pm}4.7\;vs.\;5.9{\pm}1.9$, p<0.05). In conclusion, biopsy of mouse embryos after ZD with ATS is a safe and highly efficient method for PGD, and coculture with Vero cells showed a positive effect on the development in vitro of biopsied mouse embryos and blastomeres as a preclinical model for PGD of human embryos.

• Tuberculosis and Respiratory Diseases
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• v.44 no.4
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• pp.796-805
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• 1997

Background : It is clear that deregulation of cell cycle progression is a hallmark of neoplastic transformation and genes involved in the $G_1$/S transition of the cell cycle are especially frequent targets for mutations in human cancers, including lung cancer. p16 gene product, one of the G1 cell-cycle related proteins, that is recently identified plays an important role in the negative regulation of the the kinase activity of the cyclin dependent kinase (cdk) enzymes. Therefore p16 gene is known to be an important tumor suppressor gene and is also called MTS1 (multiple tumor suppressor 1). No more oncogenes have been reported to be frequently related to multiple different malignancies than the alterations of p16 gene. Especially when it comes to non-small cell lung cancer, there was no expression of p16 in more than 70% of cell lines examined. And also it is speculated that p16 gene could exert a key role in the development of non-small cell lung cancer. This study was designed to evaluate whether p16 gene could be used as a candidate for gene therapy of non-small cell lung cancer. Methods : After the extraction of total RNA from normal fibroblast cell line and subsequent reverse transcriptase reaction and polymerase chain reaction, the amplified p16 cDNA was subcloned into eukaryotic expression plasmid vector, pRC-CMV. The constructed pRC-CMV-p16 was transfected into the NCI-H441 NSCLC cell line using lipofectin. The changes of G1 cell-cycle related proteins were investigated with Western blot analysis and immunoprecipitation after extraction of proteins from cell lysates and tumor suppressive effect was observed by clonogenic assay. Results : (1) p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 showed the formation of p16 : cdk 4 complex and decreased phosphorylated Rb protein, while control cell line did not. (2) Clonogenic assay demonstrated that the number of colony formation was markedly decreased in p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 than the control cell line. Conclusion : It is confirmed that the expression of p16 protein in p16 absent NSCLC cell line with the gene transfection leads to p16 : cdk4 complex formation, subsequent decrease of phosphorylated pRb protein and ultimately tumor suppressive effects. And also it provides the foundation for the application of p16 gene as a important candidate for the gene therapy of NSCLC.

• Transactions of the Korean Society of Mechanical Engineers B
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• v.38 no.6
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• pp.493-500
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• 2014

Nonlinear characteristics of cellular counterflow diffusion flame near the radiative extinction limit at large Damk$\ddot{o}$hler number are numerically investigated. Lewis number is assumed to be 0.5 and flame evolution is calculated by imposing an infinitesimal disturbance to a one-dimensional(1-D) steady state flame. The early stage of nonlinear development is very similar to that predicted in a linear stability analysis. The disturbance with the wavenumber of the fastest growing mode emerges and grows gradually. Eventual, an alternating pattern of reacting and quenching stripes is developed. The cellular flame temperature is higher than that of 1-D flame because of the gain of the total enthalpy. As the Damk$\ddot{o}$hler number is further increased, the shape of the cell becomes circular to increase the surface area per unit reacting volume. The cellular flames do not extinguish but survive even above the 1-D steady state extinction condition.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.6
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• pp.585-590
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• 2009

The purpose of this study was to examine the effects of oral-administered sex hormone for hybrid sturgeon, bester juvenile. The bester juveniles (2 months after hatching) were received a diet containing various doses of $17\alpha$-methyltestosterone (MT) or estradiol-$17\beta$ ($E_2$) for 6 months. Somatic growth of bester sturgeon juvenile did not show significant differences between experimental and control groups (27.9-30.5 cm; 125.1-161.7 g), although survival percentages showed a decreasing tendency in MT-treated animals. By histological examination, germ cells were recorded as smooth type in MT-treated fish and uneven type of germinal epithelium in $E_2$-treated animals. Their sex ratios were 5:4:1 (male: female: undifferentiation) in control and low dose of MT-treated fish (1 mg/kg), and 9:1:0 in fish treated with high dose of MT (10 mg/kg), whereas the ratios were reversed by both low and high doses of $E_2$ treatment, recorded as 2:8:0. Gonadal areas were not significantly differed in all trials (424,600.4 - 1,039,656.3 ${\mu}m^2$). Total number of germ cells, number of germ cells per gonadal areas and number of germ cells per area were significantly higher to 144.7-148.7 cells/section, 374.0-408.5 $cells/mm^2$ and 1,599.5-1,670.9 $cells/mm^2$ in $E_2$ treatment than those of others (30.4-63.9 cells/section, 148.4-226.9 $cells/mm^2$ and 850.0-1,050.6 $cells/mm^2$), respectively. And somatic growth according to their gender was not significantly differed between male and female.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.7
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• pp.2675-2677
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• 2015

Background: Depending on various pathological factors, non muscle invasive bladder cancer (NMIBC) shows varying degrees of recurrence. The aim of this study was to determine the incidence of recurrence of NMIBS in our centre, study the influence of intrinsic tumour characteristics like grade, stage, size and number, and compare our results with data in the published literature. Materials and Methods: A hospital based retrospective study was conducted on patients who underwent treatment for NMIBC from 2011 to 2014. The factors studied were number, size, grade, stage and site for correlation with recurrence. Statistical analysis was performed using Medcalc version 12, using Pearson's Chi square test to ascertain associations between variables. Results: A total of 73 patients with NMIBC were studied of which 48 (65.8%) had low grade and 25 (34.2%) had high grade tumours. Some 38 patients (52.1%) had Ta tumours, 34 (46.6%) had T1 and one had CIS. Mean follow up was 34.3 months. Recurrence rates were found to be 33.3% in low grade and 52.0% in high grade tumours. The overall recurrence rate in our centre was 39.7%. Significant correlations were seen between stage and recurrence, with a rate of 15% for Ta and 63.3% for T1 tumours. Fourteen out of 21 bladder cancers (66.6%) with multiple tumours demonstrated recurrence (p=0.006). Grade, size and site had no influence. Conclusions: In our study, recurrence of NMIBC was found to be directly proportional to stage and number of primary tumours, but not grade, size and site. The incidence of recurrence of NMIBC both stage wise and grade wise in our centre was also low compared to the data in the published literature.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.4
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• pp.585-592
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• 2017

Objective: The present study investigates pre- and post-implantation developmental competence of nuclear-transferred porcine embryos derived from male and female fetal fibroblasts. Methods: Male and female fetal fibroblasts were transferred to in vitro-matured enucleated oocytes and in vitro and in vivo developmental competence of reconstructed embryos was investigated. And, a total of 6,789 female fibroblast nuclear-transferred embryos were surgically transferred into 41 surrogate gilts and 4,746 male fibroblast nuclear-transferred embryos were surgically transferred into 25 surrogate gilts. Results: The competence to develop into blastocysts was not significantly different between the sexes. The mean cell number of female and male cloned blastocysts obtained by in vivo culture ($143.8{\pm}10.5$ to $159.2{\pm}14.8$) was higher than that of in vitro culture of somatic cell nuclear transfer (SCNT) groups ($31.4{\pm}8.3$ to $33.4{\pm}11.1$). After embryo transfer, 5 pregnant gilts from each treatment delivered 15 female and 22 male piglets. The average birth weight of the cloned piglets, gestation length, and the postnatal survival rates were not significantly different (p<0.05) between sexes. Conclusion: The present study found that the sex difference of the nuclear donor does not affect the developmental rate of porcine SCNT embryos. Furthermore, postnatal survivability of the cloned piglets was not affected by the sex of the donor cell.