Characterization of Tailoring Genes Involved in the Modification of Geldanamycin Polyketide in Streptomyces hygroscopicus JCM4427

( Jin Chul Shin ),( Zhu Na ),( Dong Ho Lee ),( Won Cheol Kim ),( Kyeong Lee ),( Yue Mao Shen ),( Sang Gi Paik ),( Young Soo Hong ),( Jung Joon Lee ) 한국미생물 · 생명공학회 2008 Journal of microbiology and biotechnology Vol.18 No.6

Suppression of Cellular Apoptosis Susceptibility (CSE1L) Inhibits Proliferation and Induces Apoptosis in Colorectal Cancer Cells

Zhu, Jin-Hui,Hong, De-Fei,Song, Yong-Mao,Sun, Li-Feng,Wang, Zhi-Fei,Wang, Jian-Wei Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.2

The cellular apoptosis susceptibility (CSE1L) gene has been demonstrated to regulate multiple cellular mechanisms including the mitotic spindle check point as well as proliferation and apoptosis. However, the importance of CSE1L in human colon cancer is largely unknown. In the present study, we examined expression levels of CSE1L mRNA by semiquantitative RT-PCR. A lentivirus-mediated small interfering RNA (siRNA) was used to knock down CSE1L expression in the human colon cancer cell line RKO. Changes in CSE1L target gene expression were determined by RT-PCR. Cell proliferation was examined by a high content screening assay. In vitro tumorigenesis was measured by colony-formation assay. Cell cycle distribution and apoptosis were detected by flow cytometric analysis. We found CSE1L mRNA to be expressed in human colon cancer cells. Using a lentivirus based RNAi approach, CSE1L expression was significantly inhibited in RKO cells, causing cell cycle arrest in the G2/M and S phases and a delay in cell proliferation, as well as induction of apoptosis and an inhibition of colony growth capacity. Collectively, the results suggest that silencing of CSE1L may be a potential therapeutic approach for colon cancer.

Predictive Value of the Surface-Enhanced Resonance Raman Scattering-Based MTT Assay: A Rapid and Ultrasensitive Method for Cell Viability in Situ

Mao, Zhu,Liu, Zhuo,Chen, Lei,Yang, Jin,Zhao, Bing,Jung, Young Mee,Wang, Xu,Zhao, Chun American Chemical Society 2013 ANALYTICAL CHEMISTRY - Vol.85 No.15

<P>SERRS (surface-enhanced resonance Raman scattering) has been used to develop and optimize a novel and quantitative MTT assay for living cell viability. This highly sensitive method derives from two factors for formazan signal enhancing: the addition of Au nanoparticles and the resonance effect by 632.8 nm of excitation. The results show that the background elements, such as excessive MTT residues, serum, and the drug, did not interfere with the detection of formazan. Moreover, the detection limit of formazan is as low as 1 ng/mL. With the use of this method to quantify metabolically viable cells, dose–response curves of treated and untreated cells with the drug were constructed on the human lung cancer cell A549. The results also show that the Raman signal generated is dependent on the degree of activation of the cells. In comparison to the traditional method, the main advantages of this method are its rapidity (30 min), high-selectivity, high-precision, and cost-effectiveness (0.1 mg/mL MTT) without time-consuming steps and any modifying or labeling procedure. This work reports on an improved research tool that may help researchers apply this method for in situ cell assays.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/ancham/2013/ancham.2013.85.issue-15/ac401254s/production/images/medium/ac-2013-01254s_0010.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/ac401254s'>ACS Electronic Supporting Info</A></P>

Triterpenoid Saponins from the Seeds of Caragana microphylla

Gui-Lin Jin,Cheng-Jian Zheng,Wen-Bo Xin,Zhu-Jun Mao,Pei-Xin Sun,Zhi-Xin Zeng,Lu-Ping Qin 대한약학회 2011 Archives of Pharmacal Research Vol.34 No.6

Two new triterpenoid saponins, namely caraganoside C (1) and caraganoside D (2), were isolated from the seeds of Caragana microphylla. Their structures were elucidated on the basis of spectroscopic analyses, including homo- and hetero-nuclear correlation NMR experiments (COSY, HSQC and HMBC). Both 1 and 2 exhibited moderate inhibitory activity against NO production in LPS-stimulated RAW264.7 cells with IC_50 values of 26.4 μM and 32.2 μM, respectively. In addition, 1 showed weak cytotoxicity against MCF-7, HL-60, HCT116, and A549 cell lines.

Evaluation of Microvascular Anastomosis Using Real-Time, Ultra–High-Resolution, Fourier Domain Doppler Optical Coherence Tomography

Huang, Yong,Tong, Dedi,Zhu, Shan,Wu, Lehao,Mao, Qi,Ibrahim, Zuhaib,Lee, W. P. Andrew,Brandacher, Gerald,Kang, Jin U. Williams & Wilkins 2015 Plastic and reconstructive surgery Vol.135 No.4

BACKGROUND:: Evolution in microsurgical techniques and tools has paved the way for supermicrosurgical anastomoses, with vessel diameters often approaching below 0.8 mm in the clinical realm and even smaller (0.2 to 0.3 mm) in murine models. Several imaging and monitoring devices have been introduced for postoperative monitoring, but intraoperative guidance, assessment, and predictability have remained limited to binocular optical microscopy and the surgeon’s experience. The authors present a high-resolution, real-time, three-dimensional imaging modality for intraoperative evaluation of luminal narrowing, thrombus formation, and flow alterations. METHODS:: An imaging modality that provides immediate, in-depth, high-resolution, three-dimensional structure view and flow information of the anastomosed site, called phase-resolved Doppler optical coherence tomography, was developed. Twenty-two mouse femoral artery anastomoses and 17 mouse venous anastomoses were performed and evaluated. Flow status, vessel inner lumen three-dimensional structure, and early thrombus detection were analyzed based on imaging results. Predictions formed correlated with actual long-term surgical outcomes. Eventually, four cases of mouse orthotopic limb transplantation were carried out, and predicted long-term patency based on imaging results was confirmed by actual results. RESULTS:: The assessments based on high-resolution three-dimensional visualization of the vessel flow status and inner lumen provided by phase-resolved Doppler optical coherence tomography show 92 percent sensitivity and 90 percent specificity for arterial anastomoses and 90 percent sensitivity and 86 percent specificity for venous anastomoses. CONCLUSIONS:: Phase-resolved Doppler optical coherence tomography is an effective evaluation tool for microvascular anastomosis. It can predict the long-term vessel patency with high sensitivity and specificity.

Production of Citrate by Anaerobic Fungi in the Presence of Co-culture Methanogens as Revealed by ¹H NMR Spectrometry

Cheng, Yan Fen,Jin, Wei,Mao, Sheng Yong,Zhu, Wei-Yun Asian Australasian Association of Animal Productio 2013 Animal Bioscience Vol.26 No.10

The metabolomic profile of the anaerobic fungus Piromyces sp. F1, isolated from the rumen of goats, and how this is affected by the presence of naturally associated methanogens, was analyzed by nuclear magnetic resonance spectroscopy. The major metabolites in the fungal monoculture were formate, lactate, ethanol, acetate, succinate, sugars/amino acids and ${\alpha}$-ketoglutarate, whereas the co-cultures of anaerobic fungi and associated methanogens produced citrate. This is the first report of citrate as a major metabolite of anaerobic fungi. Univariate analysis showed that the mean values of formate, lactate, ethanol, citrate, succinate and acetate in co-cultures were significantly higher than those in the fungal monoculture, while the mean values of glucose and ${\alpha}$-ketoglutarate were significantly reduced in co-cultures. Unsupervised principal components analysis revealed separation of metabolite profiles of the fungal mono-culture and co-cultures. In conclusion, the novel finding of citrate as one of the major metabolites of anaerobic fungi associated with methanogens may suggest a new yet to be identified pathway exists in co-culture. Anaerobic fungal metabolism was shifted by associated methanogens, indicating that anaerobic fungi are important providers of substrates for methanogens in the rumen and thus play a key role in ruminal methanogenesis.

Research on User-personalized Image Retrieval Method

Yu Song,Jing-fei Ren,Mao-Zhu Jin,Pei-Yu Ren 보안공학연구지원센터 2014 International Journal of Multimedia and Ubiquitous Vol.9 No.6

Hepatocyte growth factor and soluble cMet levels in plasma are prognostic biomarkers of mortality in patients with severe acute kidney injury

( Lilin Li ),( Jung Nam An ),( Jeonghwan Lee ),( Dong Jin Shin ),( Shi Mao Zhu ),( Jin Hyuk Kim ),( Dong Ki Kim ),( Dong-ryeol Ryu ),( Sejoong Kim ),( Jung Pyo Lee ) 대한신장학회 2021 Kidney Research and Clinical Practice Vol.40 No.4

Background: Hepatocyte growth factor (HGF)/cMet pathway is necessary for repair and regeneration following acute kidney injury (AKI). We evaluated the clinical potential of plasma HGF and soluble cMet as prognostic biomarkers for severe AKI requiring continuous renal replacement therapy (CRRT). Methods: One hundred thirty-six patients with severe AKI who participated in the VENUS (volume management under body composition monitoring in critically ill patients on CRRT) trial between 2017 and 2019 were enrolled in this study. We investigated associations between plasma HGF and cMet concentrations and all-cause mortality. Results: Plasma HGF and soluble cMet levels were positively correlated. Patients were divided into three groups based on their HGF and soluble cMet concentrations. The day D 0, D2, and D7 highest concentration HGF groups had significantly higher in-hospital mortality after adjusting for sex, body mass index, Acute Physiology and Chronic Health Evaluation II, and age-adjusted Charlson comorbidity index score, especially on D7 (hazard ratio, 4.26; 95% confidence interval, 1.71-10.62; p = 0.002). D7 soluble cMet level was also associated with mortality. Receiver operating characteristic curve analysis indicated that D7 HGF and soluble cMet levels were best at predicting mortality. Addition of plasma HGF and soluble cMet to conventional prognostic indices significantly improved the predictive value for mortality on D7. However, plasma HGF and soluble cMet were not associated with fluid status. Conclusion: Plasma HGF and soluble cMet levels were significant predictors of the outcomes of severe AKI patients undergoing CRRT. There was no correlation between plasma HGF and soluble cMet levels and fluid balance.

Establishment and evaluation of a murine anb3-integrinexpressing cell line with increased susceptibility to Foot-and-mouth disease virus

Wei Zhang,Kaiqi Lian,Fan Yang,Yang Yang,Zhijian Zhu,Zixiang Zhu,Weijun Cao,Ruoqing Mao,Ye Jin,Jijun He,Jianhong Guo,Xiangtao Liu,Haixue Zheng 대한수의학회 2015 Journal of Veterinary Science Vol.16 No.3

Integrin anb3 plays a major role in various signaling pathways, cell apoptosis, and tumor angiogenesis. To examine the functions and rolesof anb3 integrin, a stable CHO-677 cell line expressing the murine anb3 heterodimer (designated as “CHO-677-manb3” cells) wasestablished using a highly efficient lentiviral-mediated gene transfer technique. Integrin subunits an and b3 were detected at the gene andprotein levels by polymerase chain reaction (PCR) and indirect immunofluorescent assay (IFA), respectively, in the CHO-677-manb3 cellline at the 20th passage, implying that these genes were successfully introduced into the CHO-677 cells and expressed stably. A plaque-formingassay, 50% tissue culture infective dose (TCID50), real-time quantitative reverse transcription-PCR, and IFA were used to detect the replicationlevels of Foot-and-mouth disease virus (FMDV) in the CHO-677-manb3 cell line. After infection with FMDV/O/ZK/93, the cell line showeda significant increase in viral RNA and protein compared with CHO-677 cells. These findings suggest that we successfully established a stableanb3-receptor-expressing cell line with increased susceptibility to FMDV. This cell line will be very useful for further investigation of anb3integrin, and as a cell model for FMDV research.

Rational Biosynthetic Engineering for Optimization of Geldanamycin Analogues

Kim, Woncheol,Lee, Dongho,Hong, Seong Su,Na, Zhu,Shin, Jin Chul,Roh, Su Heun,Wu, Cheng-Zhu,Choi, Oksik,Lee, Kyeong,Shen, Yue-Mao,Paik, Sang-Gi,Lee, Jung Joon,Hong, Young-Soo WILEY-VCH Verlag 2009 Chembiochem Vol.10 No.7

<P>Tailor made: We report the rational biosynthesis of C15 hydroxylated non-quinone geldanamycin analogues by site-directed mutagenesis of the geldanamycin polyketide synthase (PKS), together with a combination of post-PKS tailoring genes. Rational biosynthetic engineering allowed the generation of geldanamycin derivatives, such as DHQ3 illustrated in the figure, which had superior pharmacological properties in comparison to the parent compound. <img src='wiley_img/14394227-2009-10-7-CBIC200800763-content.gif' alt='wiley_img/14394227-2009-10-7-CBIC200800763-content'> </P><P>A rational biosynthetic engineering approach was applied to the optimization of the pharmacological properties of the benzoquinone ansamycin, geldanamycin. Geldanamycin and its natural or semisynthetic derivatives have the potential to serve as anticancer chemotherapeutic agents. However, these first-generation Hsp90 inhibitors share an unfavorable structural feature that causes both reduced efficacy and toxicity during clinical evaluation. We report the rationally designed biosynthesis of C15 hydroxylated non-quinone geldanamycin analogues by site-directed mutagenesis of the geldanamycin polyketide synthase (PKS), together with a combination of post-PKS tailoring genes. A 15-hydroxyl-17-demethoxy non-quinone analogue, DHQ3, exhibited stronger inhibition of Hsp90 ATPase activity (4.6-fold) than geldanamycin. Taken together, the results of the present study indicate that rational biosynthetic engineering allows the generation of derivatives of geldanamycin with superior pharmacological properties.</P> <B>Graphic Abstract</B> <P>Tailor made: We report the rational biosynthesis of C15 hydroxylated non-quinone geldanamycin analogues by site-directed mutagenesis of the geldanamycin polyketide synthase (PKS), together with a combination of post-PKS tailoring genes. Rational biosynthetic engineering allowed the generation of geldanamycin derivatives, such as DHQ3 illustrated in the figure, which had superior pharmacological properties in comparison to the parent compound. <img src='wiley_img/14394227-2009-10-7-CBIC200800763-content.gif' alt='wiley_img/14394227-2009-10-7-CBIC200800763-content'> </P>