This study was conducted to investigate the effects of donor cell treatments on the production of transgenic cloned piglets. Ear fibroblast cell obtained from NIH MHC Inbred minipig was used as control. The GalT knock-out/CD45 knock-in (GalT/CD46) transgenic cell lines were established and used as donor cells. The reconstructed GalT/CD46 embryos were surgically transferred into oviduct of naturally cycling surrogate sows (Landrace ${\times}$ Yorkshire) on the second day of standing estrus. Unlike control (1.2 kV/cm, 75.4%), the fusion rate of the GalT/CDl6 donor cells was significantly higher in 1.5 kV/cm, (84.5%) than that of 1.25 kV/cm, (20.2%) (p<0.01). When the number of the transferred embryos were more than 129, the pregnancy and delivery rates were increased to 13/20 (65%) and 5/20 (25%) compared to less then 100 group [1/6 (16.7%) and 0/6 (0%)], respectively. To analyze the effect of donor cell culture condition on pregnancy and delivery rates, the GalT/CD46 donor cells were cultured with DMEM or serum reduced medium. In serum reduced medium group, the pregnancy and delivery rates were improved to 8/12 (66.7%) and 5/12 (41.7%) compared to DMEM group [3/7 (42.9%) and 0/7 (0%)], respectively. In conclusion, it can be postulated that an appropriate fusion condition and culture system is essential factors to increase the efficiency of the production of transgenic cloned piglets.
Kim, Jun-Sang;Lee, Sheng-Jin;Kim, Jin-Man;Cho, Moon-June
Radiation Oncology Journal
/
v.26
no.1
/
pp.56-64
/
2008
Purpose: Cathepsin D(CD) is a lysosomal acid proteinase that is related to malignant progression, invasion, and a poor prognosis in several tumors. The aim of this study was to evaluate the prognostic clinical significance of CD and p53 expression in pretreatment biopsy specimens from patients with locally advanced rectal cancer who were treated with preoperative chemoradiation. Materials and Methods: Eighty-nine patients with locally advanced rectal cancer(cT3/T4 or N+) were included in this study. Preoperative chemoradiation consisted of a dose of 50.4 Gy of pelvic radiation and two concurrent cycles of administration of 5-fluorouracil and leucovorin. Surgery was performed six weeks after chemoradiation. CD and p53 expression in pretreatment formalin-fixed paraffin-embedded tumor biopsy specimens were assessed by immunohistochemical staining using a CD and p53 monoclonal antibodies. The threshold value for a positive stain in tumor tissue and stromal cells was 1+ intensity in 10% of the tumors or stromal cells, respectively. Results: Positive CD expression was found in 57(64%) of the tumors and 32(35%) of the stromal cell specimens. There was no association with CD expression of the tumor or stromal cells and patient characteristics. There was a correlation between tumor CD expression with stromal cell CD expression(p=0.01). Overexpression of p53 was not a significant prognostic factor. The 5-year overall survival(OS) and disease-free survival(DFS) rates were not different between tumor CD-negative and positive patient biopsy samples(69% vs. 65%, 60% vs. 61%, respectively). The 5-year OS rates in the tumor-negative/stromal cell-negative, tumor-negative/stromal cell-positive, tumor-positive/stromal cell-negative and tumor-positive/stromal cell-positive biopsy samples were 75%, 28%, 62%, and 73%, respectively. Stromal cell staining only without positive tumor staining demonstrated the worst overall survival prognosis for patients(p=0.013). Conclusion: Overexpression of p53 in rectal biopy tissue was not associated with prognostic significance. In the pretreatment biopsy specimens, an exclusive increase in CD expression in stromal cells without tumor expression was related to poor overall survival in patients with locally advanced rectal cancer treated with preoperative chemoradiation.
Park, Hyo-Jin;Kim, Ji-Yeon;Jung, Kyung-In;Kim, Tae-Jin
IMMUNE NETWORK
/
v.9
no.4
/
pp.138-146
/
2009
Background: The MHC region of the chromosome contains a lot of genes involved in immune responses. Here we have investigated the mouse NG29/Cd320 gene in the centrometrically extended MHC region of chromosome 17. Methods: We cloned the NG29 gene by RT-PCR and confirmed the tissue distribution of its gene expression by northern blot hybridization. We generated the NG29 gene expression constructs and polyclonal antibody against the NG29 protein to perform the immunofluorescence, immunoprecipitation and flow cytometric analysis. Results: The murine NG29 gene and its human homologue, the CD320/8D6 gene, were similar in the gene structure and tissue expression patterns. We cloned the NG29 gene and confirmed its expression in plasma membrane and intracellular compartments by transfecting its expresssion constructs into HEK 293T cells. The immunoprecipitation studies with rabbit polyclonal antibody raised against the NG29-NusA fusion protein indicated that NG29 protein was a glycoprotein of about 45 kDa size. A flow cytometric analysis also showed the NG29 expression on the surface of Raw 264.7 macrophage cell line. Conclusion: These findings suggested that NG29 gene in mouse extended MHC class II region was the orthologue of human CD320 gene even though human CD320/8D6 gene was located in non-MHC region, chromosome 19p13.
This study investigated the effects of the application of a variety of ICT (Information communication Technology) cause the effects on self-directed learning characteristics, creativity and science achievement. In order to achieve the above aim, WBI (Web-Based Instructions), instructions using CD-ROM Titles and Instruction Module Using ICT for the unit were applied and analyzed which belongs to the area of 'the earth' in the subject 'science' for Elementary school students. Instructions using WBI materials, CD-ROM titles and Instruction Module Using ICT were found to be effective on 'science achievement', 'self-directed learning characteristics' and 'creativity'. They show meaningful effect on improving 'science achievement', 'self-directed learning characteristics' and 'creativity'. WBI proved more fruitful than instruction using CO-ROM titles, Instruction Module Using ICT on improving self-directed learning characteristics. Instruction Module Using ICT proved more fruitful than instruction using CD-ROM title, WBI on improving creativity.
Although there are many reports on the splenic (systemic) T cell response after Toxoptasma gondii infection, little information is available regarding the local T cell responses of peritoneal exudate cells (PEC) and gut intraepithelial Iymphocytes (IEL) following peroral infection with bradyzoites. Mice were infected with 40 cysts of the 76K strain of T. gondii, and then sacrificed at days 0, 1, 4, 7 and 10 postinfection (PI). The cellular composition and T cell responses of PEC and IEL were analyzed. The total number of PEC and IEL per mouse increased after infection, but the ratio of increase was higher in IEL. Lymphocytes were the major component of both PEC and IEL. The relative percentages of PEC macrophages and neutrophils/eosinophils increased signiflcantly at day 1 and 4 PI, whereas those of IEL did not change significantly. The percentage of PEC NK1.1 and ${\gamma\delta}T$ cells peaked at day 4 PI (p < 0.0001), and CD4 and $CD8{\alpha}T$ cells increased continuously after infection. The percentages of IEL $CD8{\alpha}$ and ${\gamma\delta}T$ cells decreased slightly at first, and then increased. CD4 and NK1.1 T cells of IEL did not change significantly after infection. $IFN-{\gamma}-producing$ PEC NK1.1 T cells increased significantly from day 1 PI, but the other T cell subsets produced $IFN-{\gamma}$ abundantly thereafter. The proportion of IEL $IFN-{\gamma}-producing$$CD8{\alpha}$ and ${\gamma\delta}T$ cells increased significantly after infection, while IEL NK1.1 T cells had similar $IFN-{\gamma}$ production patterns. Taken together, CD4 T cells were the major phenotype and the important $IFN-{\gamma}$ producing T cell subsets in PEC after oral infection with T. gondii whereas $CD8{\alpha}T$ cells had these roles in IEL. These results suggest that PEC and IEL comprise different cell differentials and T cell responses, and according to infection route these factors may contribute to the different cellular immune responses.
The purpose of this study was to investigate the effect of dietary sea-tangle extracts on blood glucose levels, serum lipid levels, thiobarbituric acid reactive substance (TBARS) and glutathione enzymes in diabetic rats treated with streptozotocin (STZ) Four groups of rats (Sprague-Dawley male rats, 180 - 200g) were consisted of normal rats fed control diet (C), diabetic rats fed control diet (CD), normal rats fed sea-tangl extracts diet (E), and diabetic rats fed sea-tangle extracts diet (ED). Diabetes was induced by single injection of streptozotocin (60 mg/kg B.W.). After 7 weeks, rats were sacrificed, serum glucose, serum total cholesterol, triglyceride levels and glutathione enzymes were measured. Urine was significantly higher in CD and ED groups than those of others (p < 0.05). Levels of amylase, calcium, uric acid, hemoglobin, cholesterol and low density lipoprotein (LDL)-cholesterol were different among four groups. But high density cholesterol (HDL)-cholesterol of ED group was significantly higher (p < 0.05) than other groups (C and E group) And the weekly change of serum glucose was decreased in the 3th,4th and 5th weeks. But serum triglyceride (TG) of diabetic rats fed sea-tangle extracts diet (ED) was lower than diabetic rats fed control diet (CD). Activity of hepatic microsomal G6Pase was significantly increased CD and ED groups higher than C and E group, but kidney was decreased ED group. Hepateic glutathione S-transferase (GST) of CD and ED group were significantly lower than C and E group (p<0.05), glutathione peroxidase (GPX) of E and ED group were significantly higher than C and CD group (p<0.05), glutathione reductase (GR) activities of ED group was significantly lower than other groups, malondialdehyde (MDA) of ED was lower than E and CD group, but kidney was increased significant in ED group compared to liver. These results suggested that dietary sea-tangle extracts reduce .hepatic disorders such as oxidant than kidney. In conclusion, dietary sea-tangle extracts groups reduced blood TG and hepatic MDA levels in STZ-induced diabetic rats.
Objectives : The aim of this study was to investigate the effect of Peucedani Radix herbal acupuncture(PR-HA) at St36(joksamni) on ovalbumin-induced asthma in mice. Methods : C57BL/6 mice were sensitized and challenged with OVA(ovalbumin) for 12 weeks(once a week) C57BL/6 mice were injected, inhaled and sprayed with OVA for 12 weeks (3times a week). One of the two experimental groups was just treated with needle-prick on St36 and the other group was treated with 1% concentrations of PR-HAS at St36, for the later 8 weeks (3times /week). Results : 1. The weight and total cells of lung of the mice group treated with PR-HA decreased significantly compared with those of control group. 2. Total Leukocytes and Eosinophils in BALF of the mice group treated with PR-HA decreased significantly compared with those of control group. 3. Eosinophils in BALF of the mice group treated with PR-HA in Photomicrographs decreased significantly compared with those of control group. 4. According to Histological analysis of lung sections, it decreased significantly adhension of collagen in PR-HA than those of control group 5. The concentration of IgE, IL-4, IL-5, in BALF and IL-4, IL-5, Il-13 in serum of the mice group treated with PR-HA decreased significantly compared with that of control group. 6. The number of Gr-1+/CD11b+, CD11b+, CD3-/CCR3+, CD4+, CD3e+/CD69+ , CD23+B220+ cells in the lungs of the mice group treated with PR-HA decreased significantly compared with those of control group. 7. The cytokine's manifestation of mRNA of the mice group treated with PR-HA with RT-PCR decreased significantly compared with that of control group. Conclusion : We conclude that PR-HA is effective on OVA-induced asthma of C57BL/6 mouse.
Park, Su-Hyung;Chang, Jun;Yang, Se-Hwan;Kim, Hye-Ju;Kwak, Hyun-Hee;Kim, Byong-Moon;Lee, Sung-Hee;Sung, Young-Chul
IMMUNE NETWORK
/
v.7
no.4
/
pp.186-196
/
2007
Background: Although IL-12 has been widely accepted to playa central role in the control of pathogen infection, the use of recombinant IL-12 (rIL-12) as a vaccine adjuvant has been known to be ineffective because of its rapid clearance in the body. Methods: To investigate the effect of sustained release of IL-12 in vivo in the peptide and protein vaccination models, rIL-12 was encapsulated into poly ($A_{DL}$-lactic-co-glycolic acid) (PLGA). Results: We found that codelivery of IL-12-encapsulated microspheres (IL-12EM) could dramatically increase not only antibody responses, but also antigen-specific $CD4^+\;and\;CD8^+$ T cell responses. Enhanced immune responses were shown to be correlated with protective immunity against influenza and respiratory syncytial virus (RSV) virus challenge. Interestingly, the enhancement of $CD8^+$ T cell response was not detectable when $CD4^+$ T cell knockout mice were subjected to vaccination, indicating that the enhancement of the $CD8^+$ T cell response by IL-12EM is dependent on $CD4^+$ T cell "help". Conclusion: Thus, IL-12EM could be applied as an adjuvant of protein and peptide vaccines to enhance protective immunity against virus infection.
Human activities have enhanced the influx of heavy metals to aquatic ecosystem and hanged the abiotic environment such as the sediments supporting benthic organisms. The levels of heavy metals (Cd, Cr, Cu, Ni, Pb and Zn) in the benthic gastropod Oxyloma hirasei and the sediments around their habitat were investigated to examine heavy metal levels and the potential of the gastropod as a bioindicator. We detected different levels of heavy metals in the sediments of two wetland areas, Upo and Mokpo, in the relatively well-conserved Upo wetland, Ramsar Convention Area. Oxyloma hirasei had higher concentrations of heavy metals except Cr and Ni in their soft tissues than in their shells (Cd: $2.10{\sim}3.16$, Cu: $19.73{\sim}28.66$, Pb: $0.67{\sim}1.17$, Zn: $216.1{\sim}285.7\;{\mu}g/g$ dry weight in the soft tissues; Cr: $1.19{\sim}2.58$, Ni: $0.47{\sim}1.16\;{\mu}g/g$ dry weight in the shells). Differences in the Cd, Cu, Ni and Pb concentrations in O. hirasei soft tissues reflected differences in heavy metal concentrations in the sediments at the sampling sites. The coefficients of variation for Cd, Cu and Pb were lower than those for other metals in the soft tissues. Levels of Cd in the tissues of O. hirasei were the highest among the metals examined in this study. Therefore, the soft tissue of O. hirasei appears to be a promising bioindicator particularly for Cd.
Herbal medicine has a high body absorption rate when it ferments. Biological and clinical research on the fermented herb gradually increases because it has effective materials for the treatment of a disease and it is a little bitter. In this study, we investigated the effect of fermented Baedokhwanbalhyobang (BDHBH) on attenuation of the development of atopic dermatitis in NC/Nga mice by evaluating the cytokine level in serum, the mRNA expression of cytokine and histological alteration of the skin, and the skin severity. We have come to the following conclusion. BDHBH led to a significant decrease in the skin severity score (63.1%) as compared to the control group. CD4+/CD45+, CD4+, B220+/CD23+, and CD11b+/Gr-1+cells of peripheral mononuclear cells (PBMCs) in the BDHBH-treated group were decreased to 6.7%, 31.1%, 22.4%, 36.6%, respectively. CD3+and CD11b+/Gr-1+immune cells in dorsal skin of the BDHBH-treated group were decreased to 52.9% and 28.0%. The levels of IL-5 and IL-13 in serum of the BDHBH-treated group were inhibited to 18.8% and 5.1%. The mRNA expressions of IL-5 and IL-13 in dorsal skin were also decreased to 30.6% and 27.8% after the treatment of BDHBH. BDHBH inhibited the proliferation and differentiation of eosinophils. In histological examination, BDHBH decreased the thickness of epidermis and dermis, and infilatration of mast cells as compared to the control group. These results indicate that BDHBH inhibits the pathogenic development of atopic dermatitis in NC/Nga mice. These results may indicate that BDHBH attenuates the development of atopic dermatitis-like lesions by lowering immune cells and inflammatory cytokine levels, and that it is valuable in drug development for the treatment of atopic dermatitis. Further experiments on the components of BDHBH will be needed to better understand the effect of a fermented herb as compared to a herb.
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