In Vitro Develq:>ment and Mitochondrial Fate of Porcine-Mouse Cloned Embryos

Yan Shi Quan,Kenji Naruse,Baek Chul Kim,Su Min Choi,Myung Youn Kim,Chang Sik Park,Dong Il Jin 한국동물생명공학회(구 한국동물번식학회) 2008 Reproductive & developmental biology Vol.32 No.2

Study on track dynamic forces due to rail short-wavelength dip defects using rail vehicle-track dynamics simulations

Yan Quan Sun,Colin Cole,Maksym Spiryagin 대한기계학회 2013 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.27 No.3

A rail vehicle-track interaction dynamics model has been applied to determine the track vertical dynamic forces due to rail short-wavelength dip defects such as squat, dip joints and welds, etc., which are required in both rail vehicle acceptance procedure and track maintenance. The model is validated using the field measurement data of rail squat defects and accelerations on a vehicle axlebox. The simulated track dynamic forces – the P2 forces due to rail dipped joints have been compared with those calculated by using a well-known formula. The results are compared and the formula’s limitations have been discussed. The dependence of the track vertical dynamic forces on the rail dip defect size and vehicle speed has also been investigated.

Effects of Insulin, Transferrin and Selenium(ITS) on In Vitro Development of Porcine Parthenogenetic and Nuclear Transfer Embryos

Yan Shi Quan,Kenji Naruse,Baek Chul Kim,Hong Rye Kim,Rong Xun Han,Su Min Choi,Chang Sik Park,Dong Il Jin 한국동물생명공학회(구 한국동물번식학회) 2007 Reproductive & developmental biology Vol.31 No.4

Insulin, transferrin and selenium (ITS) complex is reported to improve in vitro development of oocytes and embryos. This study was carried out to investigate the effects of ITS during in vitro culture (IVC) of porcine parthenogenetic and nuclear transfer (NT) embryos on subsequent developmental capacity in vitro. The electrically activated oocytes were cultured in Porcine Zygote Medium (PZM-3) with various concentrations (0, 0.1, 0.5, and 1.0%) of ITS for 7 days. Also, the electrically activated reconstructed embryos were cultured in PZM-3 with various concentrations (0, 0.1, 0.5, and 1.0%) of ITS for 6 days. Addition of ITS to culture medium did not affect development of porcine parthenogenetic embryos in vitro. To test the effect of ITS on the in vitro development of porcine NT embryos, factorial experiments were also performed for in vitro maturation (IVM) medium (TCM-199) with or without 1% ITS and culture medium (PZM-3) with or without 0.5% ITS. Addition of 0.5% ITS to culture medium increased (p<0.05) the proportion of NT blastocysts compared with non-treated group. In contrast, addition of 1% ITS to culture medium was ineffective or had a detrimental effect. Also, addition of ITS only to maturation medium increased (p<0.05) the percentage of NT blastocysts formation compared with the control group. In conclusion, addition of ITS to IVM or IVC medium could improve subsequent blastocyst development of porcine NT embryos.

In Vitro Development of Interspecies Nuclear Transfer Embryos using Porcine Oocytes with Goat and Rabbit Somatic Cells

Yan Shi Quan,Kenji Naruse,Su Min Choi,Myung Youn Kim,Rong Xun Han,Chang Sik Park,Dong Il Jin 한국동물생명공학회(구 한국동물번식학회) 2008 Reproductive & developmental biology Vol.32 No.4

Interspecies somatic cell nuclear transfer (iSCNT) is a valuable tool for studying the interactions between an oocyte and somatic nucleus. The object of this study was to investigate the developmental competence of in vitro‐matured porcine oocytes after transfer of the somatic cell nuclei of 2 different species (goat and rabbit). Porcine cumulus oocytes were obtained from the follicles of ovaries and matured in TCM‐199. The reconstructed embryos were electrically fused with 2 DC pulses of 1.1 kV/cm for 30 μs in 0.3 M mannitol medium. The activated cloned embryos were cultured in porcine zygote medium‐3 (PZM‐3), mSOF or RDH medium for 7 days. The blastocyst formation rate of the embryos reconstructed from goat or rabbit fetal fibroblasts was significantly lower than that of the embryos reconstructed from porcine fetal fibroblast cells. However, a significantly higher number of embryos reconstructed from goat or rabbit fetal fibroblasts cultured in mSOF or RDH, respectively, developed to the morular stage than those cultured in PZM‐3. These results suggest that goat and bovine fetal fibroblasts were less efficacious than porcine‐porcine cloned embryos and that culture condition could be an important factor in iSCNT. The lower developmental potential of goat‐porcine and porcine‐bovine cloned embryos may be due to incompatibility between the porcine oocyte cytoplasm and goat and bovine somatic nuclei.

Korean red ginseng extract alleviates advanced glycation end product-mediated renal injury

Quan, Hai Yan,Kim, Do Yeon,Chung, Sung Hyun The Korean Society of Ginseng 2013 Journal of Ginseng Research Vol.37 No.2

The effect of Korean red ginseng (KRG) on diabetic renal damage was investigated using streptozotocin (STZ)-induced diabetic rats. The diabetic rats showed loss of body weight gain, and increases in kidney weight and urine volume, whereas the oral administration of KRG at a dose of 100 or 250 mg/kg of body weight per day for 28 d prevented these diabetes-induced physiological abnormalities. Among the kidney function parameters, elevated plasma levels of urea nitrogen and creatinine in diabetic control rats tended to be lowered in KRG-treated rats. In addition, administration of KRG at a dose of 100 mg/kg body weight in the diabetic rats showed significant decreases in serum glucose and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), implying that KRG might prevent the pathogenesis of diabetic complications caused by impaired glucose metabolism and oxidative stress. KRG also significantly reduced advanced glycation end product (AGE) formation and secretion from kidney of diabetic rats. Furthermore, KRG decreased the levels of N-(carboxymethyl) lysine and expression of AGE receptor. KRG also reduced the overexpression of cyclooxygenase-2 and inducible nitric oxide synthase in the kidney via deactivation of nuclear factor-kappa B. We also found that KRG prevented STZ-induced destruction of glomerular structure and significantly suppressed high glucose-induced fibronectin production. Taken together, KRG ameliorates abnormalities associated with diabetic nephropathy through suppression of inflammatory pathways activated by TNF-${\alpha}$ and AGEs. These findings indicate that KRG has a beneficial effect on pathological conditions associated with diabetic nephropathy.

Fusion Expression and Immunogenicity of EHEC EspA-Stx2A1 Protein: Implications for the Vaccine Development

Yan Cheng,Youjun Feng,Ping Luo,Jiang Gu,Shu Yu,Wei-jun Zhang,Yan-qing Liu,Qing-xu Wang,Quan-ming Zou,Xu-hu Mao 한국미생물학회 2009 The journal of microbiology Vol.47 No.4

Shiga toxin 2 (Stx2) is a major virulence factor for enterohemorrhagic Escherichia coli (EHEC), which is encoded by λ lysogenic phage integrated into EHEC chromosome. Stx2A1, A1 subunit of Stx2 toxin has gathered extensive concerns due to its potential of being developed into a vaccine candidate. However, the substantial progress is hampered in part for the lack of a suitable in vitro expression system. Here we report use of the prokaryotic system pET-28a::espA-Stx2A1/BL21 to carry out the fusion expression of Stx2A1 which is linked to E. coli secreted protein A (EspA) at its N-terminus. Under the IPTG induction, EspA- Stx2A1 fusion protein in the form of inclusion body was obtained successfully, whose expression level can reach about 40% of total bacterial protein at 25°C, much higher than that at 37°C. Western blot test suggested the refolded fusion protein is of excellent immuno-reactivity with both monoclonal antibodies, which are specific to EspA and Stx2A1, respectively. Anti-sera from Balb/c mice immunized with the EspA-Stx2A1 fusion protein were found to exhibit strong neutralization activity and protection capability in vitro and in vivo. These data have provided a novel feasible method to produce Stx2A1 in large scale in vitro, which is implicated for the development of multivalent subunit vaccines candidate against EHEC O157:H7 infections.

Ginsenoside Re lowers blood glucose and lipid levels via activation of AMP-activated protein kinase in HepG2 cells and high-fat diet fed mice.

Quan, Hai-Yan,Yuan, Hai-Dan,Jung, Mi Song,Ko, Sung Kwon,Park, Young Guk,Chung, Sung Hyun D.A. Spandidos 2012 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE Vol.29 No.1

<P>Ginsenoside Re is a protopanaxatriol-type saponin isolated from Panax ginseng berry. Although anti-diabetic and anti-hyperlipidemic effects of Re have been reported by several groups, its mechanism of action is largely unknown until now. Here, we examine anti-diabetic and anti-hyperlipidemic activities of Re and action mechanism(s) in human HepG2 hepatocytes and high-fat diet fed C57BL/6J mice. Re suppresses the hepatic glucose production via induction of orphan nuclear receptor small heterodimer partner (SHP), and inhibits lipogenesis via suppression of sterol regulatory element binding protein-1c (SREBP-1c) and its target gene [fatty acid synthase (FAS), stearoyl-CoA desaturase-1 (SCD1)] transcription. These effects were mediated through activation of AMP-activated protein kinase (AMPK), and abolished when HepG2 cells were treated with an AMPK inhibitor, Compound C. C57BL/6J mice were randomly divided into five groups: regular diet fed group (RD), high-fat diet fed group (HFD) and the HFD plus Re (5, 10, 20 mg/kg) groups. Re treatment groups were fed a high-fat diet for 6 weeks, and then orally administered Re once a day for 3 weeks. The in vitro results are likely to hold true in an in vivo experiment, as Re markedly lowered blood glucose and triglyceride levels and protected against hepatic steatosis in high-fat diet fed C57BL/6J mice. In conclusion, the current study suggest that ginsenoside Re improves hyperglycemia and hyperlipidemia through activation of AMPK, and confers beneficial effects on type 2 diabetic patients with insulin resistance and dyslipidemia.</P>

Licochalcone A Prevents Adipocyte Differentiation and Lipogenesis via Suppression of Peroxisome Proliferator-Activated Receptor γ and Sterol Regulatory Element-Binding Protein Pathways

Quan, Hai-Yan,Baek, Nam In,Chung, Sung Hyun American Chemical Society 2012 Journal of agricultural and food chemistry Vol.60 No.20

<P>Licochalcone A (LA) has been shown to exert multiple pharmacological effects, including anti-inflammatory, antiparasitic, antifungal, anticancer, and osteogenic activities. The present study investigated the ability of LA to suppress the differentiation of 3T3-L1 preadipocytes, and its antiobesity activity was explored using high fat diet (HFD)-fed ICR mice. During the terminal differentiation process, 3T3-L1 preadipocytes were treated with LA, and the lipid contents were quantified along with any changes in the expression of biomarkers associated with adipocyte differentiation and lipogenesis. The results show that LA significantly reduced lipid accumulation and down-regulated the expression of peroxisome proliferator-activated receptor γ, CCAAT/enhancer binding protein α, sterol regulatory element-binding protein 1c, and their target genes (fatty acid binding protein, fatty acid synthase, stearoyl-CoA desaturase 1, and glycerol-3-phosphate acyltransferase). In an animal study, body weight, triglyceride, cholesterol, and nonesterified fatty acid levels in the group given 10 mg/kg LA were significantly decreased by 14.0, 48.2, 58.9, and 73.5%, respectively. Transverse microcomputed tomography indicated that visceral fat depots in LA-treated mice were markedly reduced when compared with those of the HFD control group. In summary, these results suggest that LA exerts an antiobesity effect and that it is a candidate for future clinical trials.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jafcau/2012/jafcau.2012.60.issue-20/jf2050763/production/images/medium/jf-2011-050763_0007.gif'></P>

In Vitro Development of Interspecies Nuclear Transfer Embryos using Porcine Oocytes with Goat and Rabbit Somatic Cells

Quan, Yan Shi,Naruse, Kenji,Choi, Su-Min,Kim, Myung-Youn,Han, Rong-Xun,Park, Chang-Sik,Jin, Dong-Il The Korean Society of Animal Reproduction 2008 Reproductive & developmental biology Vol.32 No.4

Interspecies somatic cell nuclear transfer (iSCNT) is a valuable tool for studying the interactions between an oocyte and somatic nucleus. The object of this study was to investigate the developmental competence of in vitro-matured porcine oocytes after transfer of the somatic cell nuclei of 2 different species (goat and rabbit). Porcine cumulus oocytes were obtained from the follicles of ovaries and matured in TCM-199. The reconstructed embryos were electrically fused with 2 DC pulses of 1.1kV/cm for $30{\mu}s$ 0.3M mannitol medium. The activated cloned embryos were cultured in porcine zygote medium-3 (PZM-3), mSOF or RDH medium for 7 days. The blastocyst formation rate of the embryos reconstructed from goat or rabbit fetal fibroblasts was significantly lower than that of the embryos reconstructed from porcine fetal fibroblast cells. However, a significantly higher number of embryos reconstructed from goat or rabbit fetal fibroblasts cultured in mSOF or RDH, respectively, developed to the morular stage than those cultured in PZM-3. These results suggest that goat and bovine fetal fibroblasts were less efficacious than porcine-porcine cloned embryos and that culture condition could be an important factor in iSCNT. The lower developmental potential of goat-porcine and porcine-bovine cloned embryos may be due to incompatibility between the porcine oocyte cytoplasm and goat and bovine somatic nuclei.

In Vitro Develq:>ment and Mitochondrial Fate of Porcine-Mouse Cloned Embryos

Yan Shi Quan,Kenji Naruse,Baek Chul Kim,Su Min Choi,Myung Youn Kim,Chang Sik Park,Dong Il Jin 한국동물번식학회 2008 Reproductive & Developmental Biology(Supplement) Vol.32 No.2s