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Beaton, Benjamin P,Kwon, Deug-Nam,Choi, Yun-Jung,Kim, Jae-Hwan,Samuel, Melissa S,Benne, Joshua A,Wells, Kevin D,Lee, Kiho,Kim, Jin-Hoi,Prather, Randall S John WileySons, Ltd 2015 Xenotransplantation Vol.22 No.5
<P><B>Background</B></P><P>Recent advancements in gene editing techniques have increased in number and utility. These techniques are an attractive alternative to conventional gene targeting methods via homologous recombination due to the ease of use and the high efficiency of gene editing. We have previously produced cytidine monophosphate-N-acetylneuraminic acid hydroxylase (<I>CMAH</I>) knockout (KO) pigs in a Minnesota miniature pig genetic background. These pigs were generated using zinc-finger nucleases (ZFNs) in combination with donor DNA containing a total homology length of 1600 bp (800-bp homology on each arm). Our next aim was to introduce the targeted disruption of alpha-1,3-galactosyltransferase (<I>GGTA1</I>) in the <I>CMAH</I> KO genetic background and evaluate the effect of donor DNA homology length on meganuclease-mediated gene targeting.</P><P><B>Methods</B></P><P>Zinc-finger nucleases from a previous <I>CMAH</I> KO experiment were used as a proof of concept to identify a correlation between the length of donor DNA homology and targeting efficiency. Based on those results, experiments were designed to use transcription activator-like effector nucleases (TALENs) to generate bi-allelically modified <I>GGTA1</I> cells using donor DNAs carrying various lengths of homology. Donor DNA was designed to symmetrically flank the predicted cleavage sites in <I>CMAH</I> and <I>GGTA1</I> for both ZFN and TALEN cleavage sites, respectively. For both genes, the length of total homology ranged from 60 to 1799 bp. Sialyltransferase gene expression profiles were evaluated in <I>CMAH</I> and <I>GGTA1</I> double KO pig cells and were compared to wild-type and <I>CMAH</I> KO cells.</P><P><B>Results</B></P><P>Introduction of donor DNA with ZFNs demonstrated that small amounts of homology (60 bp) could facilitate homology-directed repair during ZFN-mediated targeting of <I>CMAH</I>; however, donor DNA with longer amounts of homology resulted in a higher frequency of homology-directed repair. For the <I>GGTA1</I> KO experiments that used TALENs and donor DNA, donor DNA alone did not result in detectable bi-allelic conversion of <I>GGTA1</I>. As the length of donor DNA increased, the bi-allelic disruption of <I>GGTA1</I> increased from 0.5% (TALENs alone, no donor DNA present) to a maximum of 3% (TALENs and donor DNA with total homology of 1799 bp). Inclusion of homologous donor DNA in TALEN-mediated gene targeting facilitated a higher incidence of bi-allelically modified cells. Using the generated cells, we were able to demonstrate the lack of <I>GGTA1</I> expression and the decrease in gene expression sialyltransferase-related genes.</P><P><B>Conclusions</B></P><P>The approach of using donor DNA in conjunction with a meganuclease can be used to increase the efficiency of gene targeting. The gene editing methods can be applied to other genes as well as other mammalian systems. Additionally, gene expression analysis further confirms that the <I>CMAH</I>/<I>GGTA1</I> double KO pigs can be a valuable source for the study of pig-to-human xenotransplantation.</P>
Robot-assisted distal ureteral reconstruction for benign pathology: Current state
Aeen M. Asghar,Randall A. Lee,Kevin K. Yang,Michael Metro,Daniel D. Eun 대한비뇨의학회 2020 Investigative and Clinical Urology Vol.61 No.-
Distal ureteral reconstruction for benign pathologies such as stricture disease or iatrogenic injury has posed a challenge for urologist as endoscopic procedures have poor long-term outcomes, requiring definitive open reconstruction. Over the past decade, there has been an increasing shift towards robot-assisted laparoscopy (RAL) with multiple institutions reporting their outcomes. In this article, we reviewed the current literature on RAL distal ureteral reconstruction, focusing on benign pathologies only. We present peri-operative data and outcomes on the most common technique, ureteral reimplantation, as well as adjunct procedures such as psoas hitch and Boari flap. Additionally, we present alternative techniques reported in the literature with some technical considerations. Lastly, we describe the outcomes of the comparative studies between open, laparoscopy, and RAL. Although the body of literature in this field is limited, RAL reconstruction of the distal ureter appears to be safe, feasible, and with some advantages over the traditional open approach.
Dagdeviren, Sezin,Jung, Dae Young,Lee, Eunjung,Friedline, Randall H.,Noh, Hye Lim,Kim, Jong Hun,Patel, Payal R.,Tsitsilianos, Nicholas,Tsitsilianos, Andrew V.,Tran, Duy A.,Tsougranis, George H.,Kearns American Society for Microbiology 2016 Molecular and cellular biology Vol.36 No.23
<P>Skeletal muscle insulin resistance is a major characteristic of obesity and type 2 diabetes. Although obesity-mediated inflammation is causally associated with insulin resistance, the underlying mechanism is unclear. Here, we examined the effects of chronic obesity in mice with muscle-specific overexpression of interleukin-10 (M-IL10). After 16 weeks of a high-fat diet (HFD), M-IL10 mice became markedly obese but showed improved insulin action compared to that of wild-type mice, which was largely due to increased glucose metabolism and reduced inflammation in skeletal muscle. Since leptin regulates inflammation, the beneficial effects of interleukin-10 (IL-10) were further examined in leptin-deficient ob/ob mice. Muscle-specific overexpression of IL-10 in ob/ob mice (MCK-IL10(ob/ob)) did not affect spontaneous obesity, but MCK-IL10(ob/ob) mice showed increased glucose turnover compared to that in ob/ob mice. Last, mice with muscle-specific ablation of IL-10 receptor (M-IL10R(-/-)) were generated to determine whether IL-10 signaling in skeletal muscle is involved in IL-10 effects on glucose metabolism. After an HFD, M-IL10R(-/-) mice developed insulin resistance with reduced glucose metabolism compared to that in wild-type mice. Overall, these results demonstrate IL-10 effects to attenuate obesity-mediated inflammation and improve insulin sensitivity in skeletal muscle, and our findings implicate a potential therapeutic role of anti-inflammatory cytokines in treating insulin resistance and type 2 diabetes.</P>
Thomas, Karen C,Roberts, Jessica K,Deering-Rice, Cassandra E,Romero, Erin G,Dull, Randal O,Lee, Jeewoo,Yost, Garold S,Reilly, Christopher A American Physiological Society 2012 American journal of physiology. Lung cellular and Vol.302 No.1
<P>Endogenous agonists of transient receptor potential vanilloid-1 (TRPV1) (endovanilloids) are implicated as mediators of lung injury during inflammation. This study tested the hypothesis that endovanilloids produced following lipopolysaccharide (LPS) treatment activate TRPV1 and cause endoplasmic reticulum stress/GADD153 expression in lung cells, representing a mechanistic component of lung injury. The TRPV1 agonist nonivamide induced GADD153 expression and caused cytotoxicity in immortalized and primary human bronchial, bronchiolar/alveolar, and microvascular endothelial cells, proportional to TRPV1 mRNA expression. In CF-1 mice, Trpv1 mRNA was most abundant in the alveoli, and intratracheal nonivamide treatment promoted Gadd153 expression in the alveolar region. Treatment of CF-1 mice with LPS increased Gadd153 in the lung, lactate dehydrogenase (LDH) in bronchoalveolar lavage (BAL) fluid, and lung wet-to-dry weight ratio. Cotreating mice with LPS and the TRPV1 antagonist LJO-328 reduced Gadd153 induction and LDH in BAL but did not inhibit increases in lung wet-to-dry ratio. In Trpv1(-/-) mice treated with LPS, Gadd153 induction and LDH in BAL were reduced relative to wild-type mice, and the wet-to-dry weight ratios of lungs from both wild-type and Trpv1(-/-) mice decreased. Organic extracts of blood collected from LPS-treated mice were more cytotoxic to TRPV1-overexpressing cells compared with BEAS-2B cells and extracts from control mice, however, most pure endovanilloids did not produce cytotoxicity in a characteristic TRPV1-dependent manner. Collectively, these data indicate a role for TRPV1, and endogenous TRPV1 agonists, in ER stress and cytotoxicity in lung cells but demonstrate that ER stress and cytotoxicity are not essential for pulmonary edema.</P>
BaO 과잉량에 따른 BaTiO<sub>3</sub>의 전기전도도
여홍구,국민호,김명호,송태권,배동식,박태곤,이순일,Yeo, Hong-Goo,Kuk, Min-Ho,Kim, Myong-Ho,Song, Tae-Kwon,Bae, Dong-Sik,Park, Tne-Gone,Lee, Soon-Il,Randall, Clive A. 한국세라믹학회 2005 한국세라믹학회지 Vol.42 No.5
In this study the electrical conductivity of excess BaO in $BaTiO_3$ was measured to investigate the relationship between defects and solubility in the temperature range of $900^{\circ}C$ to $1300^{\circ}C$ under various oxygen partial pressure. First of all, quenched $BaTiO_3$ powders of various Ba/Ti ratios were analysed by X-ray diffraction to confirm whether second phase is formed or not. As the results, we observed the solubility of BaO in the temperature range of $1200^{\circ}C$ to $1400^{\circ}C$, and it was also found that the conductivity minima move to lower $PO_2$ with increasing excess BaO within solubility limit.