Broad-specificity amino acid racemase, a novel non-antibiotic selectable marker for transgenic plants

Yi‑Chia Kuan,Venkatesan Thiruvengadam,Jia‑Shin Lin,Jia‑Hsin Liu,Tsan‑Jan Chen,Hsin‑Mao Wu,Wen‑Ching Wang,Liang‑Jwu Chen 한국식물생명공학회 2018 Plant biotechnology reports Vol.12 No.1

The broad-specificity amino acid racemase (Bsar) from Pseudomonas putida catalyzes the racemization of various amino acids, offering a flexible and feasible platform to develop a new non-antibiotic selectable marker system for plant transformation. In the present study, we demonstrated that a Bsar variant, Bsar-R174K, that is useful as a selectable marker gene in Arabidopsis and rice that were susceptible to l-lysine and D-alanine. The introduction of wild-type Bsar, Bsar-R174K or Bsar-R174A into E. coli lysine or asparagine auxotrophs was able to rescue the growth of these microorganisms in minimal media supplemented with selectable amino acid enantiomers. The transformation of Arabidopsis with Bsar or Bsar variants based on d-alanine selection revealed that Bsar-R174K had the greatest efficiency (2.40%), superior to kanamycin selectionbased transformation (1.10%). Whereas, l-lysine-based selection exhibited lower efficiency for Bsar-R174K (0.17%). The progenies of selected Bsar-R174K transgenic Arabidopsis revealed normal growth properties. In addition, Bsar-R174K transgenic rice was obtained on l-lysine medium with an efficiency of 0.9%, and the progenies of the transgenic rice revealed morphologically normal phenotypes comparable with their wild-type counterparts. This study presents the first report of broad range amino acid racemase Bsar-R174K as a non-antibiotic selectable marker system applied in transgenic plants.

Albumin-to-Fibrinogen Ratio as an Independent Prognostic Parameter in Untreated Chronic Lymphocytic Leukemia: A Retrospective Study of 191 Cases

Yi-Xin Zou,Jia Qiao,Hua-Yuan Zhu,Rui-Nan Lu,Yi Xia,Lei Cao,Wei Wu,Hui Jin,Wen-Jie Liu,Jin-Hua Liang,Jia-Zhu Wu,Li Wang,Lei Fan,Wei Xu,Jian-Yong Li 대한암학회 2019 Cancer Research and Treatment Vol.51 No.2

Purpose Chronic lymphocytic leukemia (CLL) is one of the most frequent type of B-cell chronic lymphoproliferative disorders and chronic inflammation takes part in the development of CLL. However, there has been no valid immune biomarker to predict the prognosis of untreated CLL patients. Materials and Methods In this retrospective study, we analyzed the clinical correlations and prognostic value of albumin-to-fibrinogen ratio (AFR) detected at diagnosis in 191 CLL patients. Results The cut-off value of AFR was 9.7 calculated by X-tile. Patients who were more than 65 years old were often accompanied by low level of AFR (p < 0.001). Survival analysis showed that patients with low level of AFR had shorter overall survival (OS) than patients with high level of AFR (p < 0.001). Multivariate analysis illustrated that AFR had a negative impact on OS (p=0.003) and was independent of parameters involved in CLL international prognostic index and other prognostic markers such as CD38 and ZAP-70. Conclusion These data provide a comprehensive view of AFR and shows that AFR at diagnosis is an adverse prognostic factor in untreated CLL patients.

Characterization of Ecdysteroid UDP-Glucosyltransferase Gene Promoter from Bombyx mori Nucleopolyhedrovirus

( Xing Jia Shen ),( Yong Zhu Yi ),( Shun Ming Tang ),( Zhi Fang Zhang ),( Yi Ren Li ),( Jia Lu He ),( Xiang Fu Wu ) 한국잠사학회 2007 International Journal of Industrial Entomology Vol.8 No.2

Bombyx mori nucleopolyhedrovirus(BmNPV) ecdysteroid UDP-glucosyltransferase gene (egt) promoter fragments of different lengths were amplified from BmNPV ZJ-8 genomic DNA by PCR. Reporter plasmids pBmegt542-luc, pBmegt309-luc and pBmegtl59-luc with luciferase (luc) driven by egt promoters were constructed. Both in vitro and in vivo expressions showed that BmNPV egt promoter activity requires the transactivation of viral factor(s), and expression of luc was detected earliest at 24 hrs post infection (pi). BmNPV ZJ-8 homologous region 3 (hr3) increased the expression of luc by over 1,600-fold. Molting hormone of 1.0∼2.0㎍/㎖ can dramatically down regulate expression of luc. Juvenile hormone analogue of 0.5∼2.0㎍/㎖ increased expression of luc by 145.8% to 75.7%. Deletion assay revealed that the promoter fragment of 159 bp contains the basal promoter structure; Promoter fragments of 309 bp and 542 bp showed similar but much higher transcriptional activities than that of 159 bp, suggesting that nucleotide from -159 to -309 nt upstream the translation initiation site harbors the main cis-acting elements.

Characterization of Ecdysteroid UDP-Glucosyltransferase Gene Promoter from Bombyx mori Nucleopolyhedrovirus

Xing-Jia Shen,Yong-Zhu Yi,Shun-Ming Tang,Zhi-Fang Zhang,Yi-Ren Li,Jia-Lu He,Xiang-Fu Wu 한국잠사학회 2004 International Journal of Industrial Entomology Vol.8 No.2

Bombyx mori nucleopolyhedrovirus (BmNPV) ecdysteroid UDP-glucosyltransferase gene (egt) promoter fragments of different lengths were amplified from BmNPV ZJ-8 genomic DNA by PCR. Reporter plasmids pBmegt542-luc, pBmegt309-luc and pBmegt159- luc with luciferase (luc) driven by egt promoters were constructed. Both in vitro and in vivo expressions showed that BmNPV egt promoter activity requires the transactivation of viral factor(s), and expression of luc was detected earliest at 24 hrs post infection (pi). BmNPV ZJ-8 homologous region 3 (hr3) increased the expression of luc by over 1,600-fold. Molting hormone of 1.0 ~ 2.0 mg/ml can dramatically down regulate expression of luc. Juvenile hormone analogue of 0.5 ~ 2.0 mg/ml increased expression of luc by 145.8% to 75.7%. Deletion assay revealed that the promoter fragment of 159 bp contains the basal promoter structure; Promoter fragments of 309 bp and 542 bp showed similar but much higher transcriptional activities than that of 159 bp, suggesting that nucleotide from -159 to -309 nt upstream the translation initiation site harbors the main cis-acting elements.

Nomogram for predicting overall survival in children with neuroblastoma based on SEER database

Song-Wu Liang,Gang Chen,Yi-Ge Luo,Peng Chen,Jin-Han Gu,Qiong-Qian Xu,Yi-Wu Dang,Li-Ting Qin,Hui-Ping Lu,Wen-Ting Huang,Zhi-Guang Huang,Li Gao,Jia-Bo Chen 대한외과학회 2020 Annals of Surgical Treatment and Research(ASRT) Vol.99 No.2

Purpose: This study was performed to establish and validate a nomogram for predicting the overall survival in children with neuroblastoma. Methods: The latest clinical data of neuroblastoma in Surveillance, Epidemiology, and End Results (SEER) database was extracted from 2000 to 2016. The cases included were randomly divided into training and validation cohorts. The survival curves were drawn with a Kaplan-Meier estimator to investigate the influences of certain single factors on overall survival. Also, least absolute shrinkage and selection operator regression was applied to further select the prognostic variables for neuroblastoma. Additionally, receiver operating characteristic (ROC) curves and calibration curves were used to evaluate the accuracy of the nomogram. Results: In total, 1,262 patients were collected and 8 independent prognostic factors were achieved, including patients’ age, sex, race, tumor grade, radiotherapy, chemotherapy, tumor site, and tumor size. Then we constructed a nomogram by using the data of the training cohort with 886 cases. Subsequently, the nomogram was validated internally and externally with 886 and 376 cases, respectively. The internal validation revealed that the area under the curves (AUC) of ROC curves of 1-, 3-, and 5-year overall survival were 0.69, 0.78, and 0.81, respectively. Accordingly, the external validation also showed that the AUC of 1-, 3-, and 5-year overall survival were all ≥0.69. Both methods of validation demonstrated that the predictive calibration curves were consistent with standard curves. Conclusion: The nomogram possess the potential to be a new tool in predicting the survival rate of neuroblastoma patients.

Intracellular Polysaccharide and its Antioxidant Activity by Pleurotus citrinopileatus SM-01

Su-Qian Wu,Shang-Long Gao,Hong-Hong Liu,Xin-Yi Sun,Long Hao,Le Jia,Li-Fei Pang,Shou-Hua Jia,Meng-Shi Jia 한국고분자학회 2013 Macromolecular Research Vol.21 No.6

The extraction parameters of intracellular polysaccharide (IPS) from Pleurotus citrinopileatus SM-01mycelia were optimized, and the in vitro and in vivo antioxidant activities of IPS were investigated. The optimum conditions of IPS extraction were predicted to be an ultrasonic treatment time of 664.09 s, precipitation time of 23.03h and pH 7.36, and IPS yield was estimated at 16.13%. The in vitro inhibition effects of IPS at a dosage of 5 g/L on the superoxide anion, 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radicals were 73.96±4.62%, 69.2±4.37%,and 50.75±4.39%, respectively, which were 72.56±5.08%, 22.83±1.94%, and 43.93±3.26% higher than that of butylated hydroxytoluene (BHT), respectively. The reducing power of IPS was 0.9±0.07, 69.81±5.24% higher than that of BHT. The activities of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT) and alanine aminotransferase (ALT) in mice blood were 241.38±23.19, 454.95±42.39, 60.32±5.16, and 32.39±2.54 U/mL,respectively, and the malonaldehyde (MDA) level was 9.54±0.72 nmol/mL. The results provided a reference for the large-scale extraction of IPS by P. citrinopileatus SM-01 in industrial fermentation, suggesting that the IPS can be used as a potential antioxidant, which enhances adaptive immune responses.

MicroRNA-146a Enhances Helicobacter pylori Induced Cell Apoptosis in Human Gastric Cancer Epithelial Cells

Wu, Kai,Yang, Liu,Li, Cong,Zhu, Chao-Hui,Wang, Xin,Yao, Yi,Jia, Yu-Jie Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.14

Helicobacter pylori (H. pylori) infection induces apoptosis in gastric epithelial cells, and this occurrence may link to gastric carcinogenesis. However, the regulatory mechanism of H. pylori-induced apoptosis is not clear. MicroRNA-146a has been implicated as a key regulator of the immune system. This report describes our discovery of molecular mechanisms of microRNA-146a regulation of apoptosis in human gastric cancer cells. We found that overexpression of microRNA-146a by transfecting microRNA-146a mimics could significantly enhance apoptosis, and this upregulation was triggered by COX-2 inhibition. Furthermore, we found that microRNA-146a density was positively correlated with apoptosis rates in H. pylori-positive gastric cancer tissues and intratumoral microRNA-146a density was negatively correlated with lymph node metastasis among H. pylori-positive gastric cancer patients. Understanding the important roles of microRNA-146a in regulating cell apoptosis in H. pylori infected human gastric cancer cells will contribute to the development of microRNA targeted therapy in the future.

Partial Least Squares Based Gene Expression Analysis in EBV-Positive and EBV-Negative Posttransplant Lymphoproliferative Disorders

Wu, Sa,Zhang, Xin,Li, Zhi-Ming,Shi, Yan-Xia,Huang, Jia-Jia,Xia, Yi,Yang, Hang,Jiang, Wen-Qi Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.11

Post-transplant lymphoproliferative disorder (PTLD) is a common complication of therapeutic immunosuppression after organ transplantation. Gene expression profile facilitates the identification of biological difference between Epstein-Barr virus (EBV) positive and negative PTLDs. Previous studies mainly implemented variance/regression analysis without considering unaccounted array specific factors. The aim of this study is to investigate the gene expression difference between EBV positive and negative PTLDs through partial least squares (PLS) based analysis. With a microarray data set from the Gene Expression Omnibus database, we performed PLS based analysis. We acquired 1188 differentially expressed genes. Pathway and Gene Ontology enrichment analysis identified significantly over-representation of dysregulated genes in immune response and cancer related biological processes. Network analysis identified three hub genes with degrees higher than 15, including CREBBP, ATXN1, and PML. Proteins encoded by CREBBP and PML have been reported to be interact with EBV before. Our findings shed light on expression distinction of EBV positive and negative PTLDs with the hope to offer theoretical support for future therapeutic study.

SDSS J013127.34-032100.1: A NEWLY DISCOVERED RADIO-LOUD QUASAR AT <i>z</i> = 5.18 WITH EXTREMELY HIGH LUMINOSITY

Yi, Wei-Min,Wang, Feige,Wu, Xue-Bing,Yang, Jinyi,Bai, Jin-Ming,Fan, Xiaohui,Brandt, William N.,Ho, Luis C.,Zuo, Wenwen,Kim, Minjin,Wang, Ran,Yang, Qian,Zhang, Ju-jia,Wang, Fang,Wang, Jian-Guo,Ai, Yanl IOP Publishing 2014 ASTROPHYSICAL JOURNAL LETTERS - Vol.795 No.2

<P>Very few of the z > 5 quasars discovered to date have been radio-loud, with radio-to-optical flux ratios (radio-loudness parameters) higher than 10. Here we report the discovery of an optically luminous radio-loud quasar, SDSS J013127.34-032100.1 (J0131-0321 in short), at z = 5.18 +/- 0.01 using the Lijiang 2.4 m and Magellan telescopes. J0131-0321 has a spectral energy distribution consistent with that of radio-loud quasars. With an i-band magnitude of 18.47 and a radio flux density of 33 mJy, its radio-loudness parameter is similar to 100. The optical and near-infrared spectra taken by Magellan enable us to estimate its bolometric luminosity to be L-bol similar to 1.1 x 10(48) erg s(-1), approximately 4.5 times greater than that of the most distant quasar known to date. The black hole mass of J0131-0321 is estimated to be 2.7 x 10(9) M-circle dot, with an uncertainty up to 0.4 dex. Detailed physical properties of this high-redshift, radio-loud, potentially super-Eddington quasar can be probed in the future with more dedicated and intensive follow-up observations using multi-wavelength facilities.</P>

Comparison of Liver Transplantation and Liver Resection for Hepatocellular Carcinoma Patients with Portal Vein Tumor Thrombus Type I and Type II

Jia-Yu Lv,Ning-Ning Zhang,Ya-Wei Du,Ying Wu,Tian-Qiang Song,Ya-Min Zhang,Yan Qu,Yu-Xin Liu,Jie Gu,Ze-Yu Wang,Yi-Bo Qiu,Bing Yang,Da-Zhi Tian,Qing-Jun Guo,Li Zhang,Ji-San Sun,Yan Xie,Zheng-Lu Wang,Xin 연세대학교의과대학 2021 Yonsei medical journal Vol.62 No.1

Purpose: The aim of this study was to compare the efficacy of liver transplantation (LT) and liver resection (LR) for hepatocellularcarcinoma (HCC) patients with portal vein tumor thrombus (PVTT) and to investigate risk factors affecting prognosis. Materials and Methods: A total of 94 HCC patients with PVTT type I (segmental PVTT) and PVTT type II (lobar PVTT) were involvedand divided into LR (n=47) and LT groups (n=47). Recurrence-free survival (RFS) and overall survival (OS) were comparedbefore and after inverse probability of treatment weighting (IPTW). Prognostic factors for RFS and OS were explored. Results: Two treatment groups were well-balanced using IPTW. In the entire cohort, LT provided a better prognosis than LR. Among patients with PVTT type I, RFS was better with LT (p=0.039); OS was not different significantly between LT and LR(p=0.093). In subgroup analysis of PVTT type I patients with α-fetoprotein (AFP) levels >200 ng/mL, LT elicited significantly longermedian RFS (18.0 months vs. 2.1 months, p=0.022) and relatively longer median OS time (23.6 months vs. 9.8 months, p=0.065). Among patients with PVTT type II, no significant differences in RFS and OS were found between LT and LR (p=0.115 and 0.335,respectively). Multivariate analyses showed treatment allocation (LR), tumor size (>5 cm), AFP and aspartate aminotransferase(AST) levels to be risk factors of RFS and treatment allocation (LR), AFP and AST as risk factors for OS. Conclusion: LT appeared to afford a better prognosis for HCC with PVTT type I than LR, especially in patients with AFP levels>200 ng/mL.