LncRNA XIST promotes carboplatin resistance of ovarian cancer through activating autophagy via targeting miR-506-3p/FOXP1 axis

Xiaoyan Xia,Zikui Li,Yaojun Li,Feng Ye,Xiaoming Zhou 대한부인종양학회 2022 Journal of Gynecologic Oncology Vol.33 No.6

Objective: Resistance to chemotherapy drugs makes ovarian cancer (OC) difficult to treat and ultimately kills patients. Long non-coding RNAs are closely related to carboplatin resistance in OC. In present study, we explored the role of lncRNA X-inactive specific transcript (XIST) on carboplatin resistance in OC. Methods: Cell viability, proliferation, and apoptosis were assessed through 2,5-diphenyl- 2H-tetrazolium bromide, colony formation, and flow cytometry assays, respectively. Microtubule-associated protein 1A/1B-light chain 3 expression was evaluated by immunofluorescence assay to analyze the cell autophagy. The interaction of XIST/ miR-506-3p or miR-506-3p/forkhead box protein P1 (FOXP1) was analyzed using RNA immunoprecipitation (RIP) and dual-luciferases reporter assays. The function of XIST/ miR-506-3p/FOXP1 axis in vivo was further confirmed by tumor xenograft study and immunohistochemistry. Results: The expression of XIST and FOXP1 increased while miR-506-3p decreased in OC and carboplatin resistance cells. XIST silencing repressed the proliferative and autophagic capacities of carboplatin resistance cells while promoted the apoptosis. XIST overexpression led to the opposite results. XIST targeted miR-506-3p and downregulated its expression. MiR-506-3p inhibition facilitated the proliferative and autophagic capacities while suppressed the apoptosis of cells, XIST knockdown reversed these effects. MiR-506-3p bound to FOXP1. XIST knockdown or miR-506-3p overexpression reversed the increase of cell proliferative and autophagic abilities and the decrease of apoptosis rate induced by FOXP1 overexpression. XIST affected autophagy and carboplatin resistance in vivo via regulating the miR-506-3p/FOXP1 axis. Conclusion: XIST knockdown inhibited autophagy and carboplatin resistance of OC through FOXP1/protein kinase B (AKT)/mammalian target of rapamycin pathway by targeting miR-506-3p.

Erythritol production by Yarrowia lipolytica mutant strain M53 generated through atmospheric and room temperature plasma mutagenesis

Xiaoyan Liu,Jinshun Lv,Jiaxing Xu,Jun Xia,Benlin Dai,Xiangqian Xu,Jiming Xu 한국식품과학회 2017 Food Science and Biotechnology Vol.26 No.4

Mutants of Yarrowia lipolytica with high erythritol production were generated through an atmospheric and room temperature plasma (ARTP) mutation system. Among these mutants, Y. lipolytica M53 exhibited the highest erythritol yield. In a batch culture, M53 produced 64.8 g/L erythritol from 100 g/L glycerol. The yields of byproducts (e.g. mannitol, arabitol, and a-ketoglutaric acid) were low, and the mechanisms underlying these changes were examined by measuring enzyme activities in the pentose phosphate pathway. Up to 145.2 g/L erythritol was produced by M53 from 200 g/L of glycerol, and erythritol accumulation was promoted by 3.7 mg/L of Cu2?, 10.15 mg/L of Mn2?, and 30.37 g/L of NaCl. Fed-batch cultivation of M53 in a 5-L fermentor produced 169.3 g/L erythritol with low levels of byproducts within 168 h. This finding confirmed the potential of M53 as an erythritol producer on a commercial scale.

vfr, A Global Regulatory Gene, is Required for Pyrrolnitrin but not for Phenazine-1-carboxylic Acid Biosynthesis in Pseudomonas chlororaphis G05

Xia Wu,Xiaoyan Chi,Yanhua Wang,Kailu Zhang,Le Kai,Qiuning He,Jinxiu Tang,Kewen Wang,Longshuo Sun,Xiuying Hao,Weihai Xie,Yihe Ge 한국식물병리학회 2019 Plant Pathology Journal Vol.35 No.4

In our previous study, pyrrolnitrin produced in Pseudomonas chlororaphis G05 plays more critical role in suppression of mycelial growth of some fungal pathogens that cause plant diseases in agriculture. Although some regulators for pyrrolnitrin biosynthesis were identified, the pyrrolnitrin regulation pathway was not fully constructed. During our screening novel regulator candidates, we obtained a white conjugant G05W02 while transposon mutagenesis was carried out between a fusion mutant G05ΔphzΔprn::lacZ and E. coli S17- 1 (pUT/mini-Tn5Kan). By cloning and sequencing of the transposon-flanking DNA fragment, we found that a vfr gene in the conjugant G05W02 was disrupted with mini-Tn5Kan. In one other previous study on P. fluorescens, however, it was reported that the deletion of the vfr caused increased production of pyrrolnitrin and other antifungal metabolites. To confirm its regulatory function, we constructed the vfr-knockout mutant G05Δvfr and G05ΔphzΔprn::lacZΔvfr. By quantifying β-galactosidase activities, we found that deletion of the vfr decreased the prn operon expression dramatically. Meanwhile, by quantifying pyrrolnitrin production in the mutant G05Δvfr, we found that deficiency of the Vfr caused decreased pyrrolnitrin production. However, production of phenazine-1-carboxylic acid was same to that in the wild-type strain G05. Taken together, Vfr is required for pyrrolnitrin but not for phenazine-1-carboxylic acid biosynthesis in P. chlororaphis G05.

Study on pharmacokinetics and tissue distribution of single dose oral tryptanthrin in Kunming mice by validated reversed-phase high-performance liquid chromatography with ultraviolet detection

Xiaoyan Zhang,Jie Xia,Wenjing Zhang,Yao Luo,Wenbo Sun,Wei Zhou 한국한의학연구원 2017 Integrative Medicine Research Vol.6 No.3

Background: Tryptanthrin is a major active constituent of several Chinese herbal plants, such as Isatidis radix. Tryptanthrin had been demonstrated to have several beneficial pharmacological effects in vitro for human diseases, including antitumor, anti-inflammatory and antibacteria activities. In contrast to the extensive in vitro investigations, the in vivo disposition process of tryptanthrin was explored limitedly. Methods: In this study, the pharmacokinetics (PK) and tissue distribution of tryptanthrin in Kunming mice following a single oral dose of 80 mg/kg tryptanthrin were investigated for the first time. Mouse plasma, liver, heart, spleen, lung, kidney and brain were collected and analyzed using a validated reversed-phase high-performance liquid chromatography with ultraviolet detection (RP-HPLC–UV) method after biological sample preparation by a simple liquid–liquid extraction. Results: The chromatographic analysis was performed on a Diamonsil C18 column (5 μm, 250 mm × 4.6 mm) and ultraviolet detection was set at a wavelength of 251 nm. The analysis was achieved with a mobile phase of methanol (A) and water (B) (60:40, v/v) at a flow rate of 1.0 mL/min. The method was linear over the concentration range of 4.0–400.0 μg/mL with a lower limit of quantification of 0.10–0.30 μg/mL. Inter- and intraday precisions (relative standard deviations %) were all within 2.93%. Recoveries of tryptanthrin were more than 86.44%. Maximal tryptanthrin concentrations in plasma and tissues of mice were reached within 2.5 hours. The actual highest concentration (Cmax) in mouse plasma was 3.13 μg/mL, the area under the curve (AUC0–t) was 9.38 h μg/mL, and the terminal half-life was 2.27 hours. The volume of distribution was 343.89 mL, the clearance rate was 204.58 mL/h, and the PK of tryptanthrin in mice after oral administration was fit to 2 compartment 1 st Order. After oral dosing of tryptanthrin to Kunming mice, the analyte was well distributed to the plasma and main tissues. Cmax was found in the liver with a mean value of 3.54 μg/g, followed by that in the kidney, lung, spleen, heart, and brain. Conclusion: In this study, a validated RP-HPLC–UV method was developed and successfully applied to PK and tissue distribution of oral tryptanthrin in mice. We confirmed that tryptanthrin was closely related and targeted to plasma, liver, kidney, and lung. These results indicate that tryptanthrin will have a good clinical application in the liver, kidney, or lung. The clinical use of tryptanthrin should focus on its pharmacodynamics and safety study in these tissues.

Genome-wide association analysis of nine reproduction and morphological traits in three goat breeds from Southern China

Sun Xiaoyan,Jiang Jing,Wang Gaofu,Zhou Peng,Li Jie,Chen Cancan,Liu Liangjia,Li Nianfu,Xia Yuanyou,Ren Hangxing 아세아·태평양축산학회 2023 Animal Bioscience Vol.36 No.2

Objective: This study aimed to investigate the significant single nucleotide polymorphisms (SNPs) and genes associated with nine reproduction and morphological traits in three breed populations of Chinese goats. Methods: The genome-wide association of nine reproduction and morphological traits (litter size, nipple number, wattle, skin color, coat color, black dorsal line, beard, beard length, and hind leg hair) were analyzed in three Chinese native goat breeds (n = 336) using an Illumina Goat SNP50 Beadchip. Results: A total of 17 genome-wide or chromosome-wide significant SNPs associated with one reproduction trait (litter size) and six morphological traits (wattle, coat color, black dorsal line, beard, beard length, and hind leg hair) were identified in three Chinese native goat breeds, and the candidate genes were annotated. The significant SNPs and corresponding putative candidate genes for each trait are as follows: two SNPs located on chromosomes 6 (CSN3) and 24 (TCF4) for litter size trait; two SNPs located on chromosome 9 (KATNA1) and 1 (UBASH3A) for wattle trait; three SNPs located on chromosome 26 (SORCS3), 24 (DYM), and 20 (PDE4D) for coat color trait; two SNPs located on chromosome 18 (TCF25) and 15 (CLMP) for black dorsal line trait; four SNPs located on chromosome 8, 2 (PAX3), 5 (PIK3C2G), and 28 (PLA2G12B and OIT3) for beard trait; one SNP located on chromosome 18 (KCNG4) for beard length trait; three SNPs located on chromosome 17 (GLRB and GRIA2), 28 (PGBD5), and 4 for hind leg hair trait. In contrast, there were no SNPs identified for nipple number and skin color. Conclusion: The significant SNPs or genes identified in this study provided novel insights into the genetic mechanism underlying important reproduction and morphological traits of three local goat breeds in Southern China as well as further potential applications for breeding goats.

vfr, A Global Regulatory Gene, is Required for Pyrrolnitrin but not for Phenazine-1-carboxylic Acid Biosynthesis in Pseudomonas chlororaphis G05

Wu, Xia,Chi, Xiaoyan,Wang, Yanhua,Zhang, Kailu,Kai, Le,He, Qiuning,Tang, Jinxiu,Wang, Kewen,Sun, Longshuo,Hao, Xiuying,Xie, Weihai,Ge, Yihe The Korean Society of Plant Pathology 2019 Plant Pathology Journal Vol.35 No.4

In our previous study, pyrrolnitrin produced in Pseudomonas chlororaphis G05 plays more critical role in suppression of mycelial growth of some fungal pathogens that cause plant diseases in agriculture. Although some regulators for pyrrolnitrin biosynthesis were identified, the pyrrolnitrin regulation pathway was not fully constructed. During our screening novel regulator candidates, we obtained a white conjugant G05W02 while transposon mutagenesis was carried out between a fusion mutant $G05{\Delta}phz{\Delta}prn::lacZ$ and E. coli S17-1 (pUT/mini-Tn5Kan). By cloning and sequencing of the transposon-flanking DNA fragment, we found that a vfr gene in the conjugant G05W02 was disrupted with mini-Tn5Kan. In one other previous study on P. fluorescens, however, it was reported that the deletion of the vfr caused increased production of pyrrolnitrin and other antifungal metabolites. To confirm its regulatory function, we constructed the vfr-knockout mutant $G05{\Delta}vfr$ and $G05{\Delta}phz{\Delta}prn::lacZ{\Delta}vfr$. By quantifying ${\beta}-galactosidase$ activities, we found that deletion of the vfr decreased the prn operon expression dramatically. Meanwhile, by quantifying pyrrolnitrin production in the mutant $G05{\Delta}vfr$, we found that deficiency of the Vfr caused decreased pyrrolnitrin production. However, production of phenazine-1-carboxylic acid was same to that in the wild-type strain G05. Taken together, Vfr is required for pyrrolnitrin but not for phenazine-1-carboxylic acid biosynthesis in P. chlororaphis G05.

Efficient Cu(OTf)2-catalyzed synthesis of novel and diverse 2,3-dihydroquinazolin-4(1H)-ones.

Zhu, Xiaoyan,Kang, So Rang,Xia, Likai,Lee, Jihye,Basavegowda, Nagaraj,Lee, Yong Rok ESCOM Science Publishers 2015 Molecular diversity Vol.19 No.1

<P>An efficient one-pot synthesis of various 2,3-dihydroquinazolin-4(1H)-one derivatives was accomplished using Cu(OTf)2-catalyzed multi-component reactions between isatoic anhydride, ketones, and amines. The method has several significant advantages; mild reaction conditions, easy handling, and efficiency of catalyst.</P>

Characterization of Carbapenemase Genes in Enterobacteriaceae Species Exhibiting Decreased Susceptibility to Carbapenems in a University Hospital in Chongqing, China

Yun Xia,Zhenzhen Liang,Xiaoyan Su,Ying Xiong 대한진단검사의학회 2012 Annals of Laboratory Medicine Vol.32 No.4

Background: Our study was to investigate the prevalence of carbapenemase genes in strains of Enterobacteriaceae species exhibiting decreased susceptibility to carbapenems in our hospital. Methods: The carbapenemase producing Enterobacteriaceae species were confirmed by modified Hodge test (MHT) and EDTA-disc synergy test which indicating the production of class B carbapenemases. PCR and sequencing analysis were used to identify the drugresistant genes. DNA fingerprinting based on enterobacterial repetitive intergenic consensus (ERIC)-PCR was applied to investigate the homology of Enterobacteriaceae species. Results: From a collection of 1,472 Enterobacteriaceae species, 18 isolates with decreased susceptibility to carbapenem treatment were identified and 9 of which were positive by MHT, and 6 of which produced class B carbapenemases. PCR and sequencing analysis of the 18 isolates revealed 4 different carbapenemase genes (blaIMP-8, blaoxa-1, blaIMP-26, and blaoxa-47) in 10 isolates, with the blaIMP-8 and blaoxa-1 genes being the most common (60-70% prevalence). ERIC-PCR showed 5, 2, and 2 unique genotypes for Enterobacter cloacae, Escherichia coli, and Klebsiella pneumoniae, respectively. Three E. coli strains isolated from different patients from the urologic surgery department exhibited the same DNA banding pattern, suggesting a possible clonal dissemination. Majority (17/18) of the carbapenem- unsusceptible Enterobacteriaceae species isolates was obtained from the surgery department of our hospital. Conclusions: The main carbapenemase genes of Enterobacteriaceae species in our hospital were blaIMP-8 and blaoxa-1. Prevalence of carbapenem resistance may be existed in surgery department and infection control should be taken for preventing further dissemination of drug-resistant strains. Background: Our study was to investigate the prevalence of carbapenemase genes in strains of Enterobacteriaceae species exhibiting decreased susceptibility to carbapenems in our hospital. Methods: The carbapenemase producing Enterobacteriaceae species were confirmed by modified Hodge test (MHT) and EDTA-disc synergy test which indicating the production of class B carbapenemases. PCR and sequencing analysis were used to identify the drugresistant genes. DNA fingerprinting based on enterobacterial repetitive intergenic consensus (ERIC)-PCR was applied to investigate the homology of Enterobacteriaceae species. Results: From a collection of 1,472 Enterobacteriaceae species, 18 isolates with decreased susceptibility to carbapenem treatment were identified and 9 of which were positive by MHT, and 6 of which produced class B carbapenemases. PCR and sequencing analysis of the 18 isolates revealed 4 different carbapenemase genes (blaIMP-8, blaoxa-1, blaIMP-26, and blaoxa-47) in 10 isolates, with the blaIMP-8 and blaoxa-1 genes being the most common (60-70% prevalence). ERIC-PCR showed 5, 2, and 2 unique genotypes for Enterobacter cloacae, Escherichia coli, and Klebsiella pneumoniae, respectively. Three E. coli strains isolated from different patients from the urologic surgery department exhibited the same DNA banding pattern, suggesting a possible clonal dissemination. Majority (17/18) of the carbapenem- unsusceptible Enterobacteriaceae species isolates was obtained from the surgery department of our hospital. Conclusions: The main carbapenemase genes of Enterobacteriaceae species in our hospital were blaIMP-8 and blaoxa-1. Prevalence of carbapenem resistance may be existed in surgery department and infection control should be taken for preventing further dissemination of drug-resistant strains.

Simple and Sensitive Electrochemical Sandwich-type Immunosensing of Human Chorionic Gonadotropin based on b-cyclodextrin Functionalized Graphene

Linfen Xu,Ling liu,Xiaoyan Zhao,Jinyu Lin,Shaohan Xu,Jinlian He,Debin Jiang,Yong Xia The Korean Electrochemical Society 2023 Journal of electrochemical science and technology Vol.14 No.1

The effective detection of human chorionic gonadotropin (HCG) is considerably important for the clinical diagnosis of both of early pregnancy and nonpregnancy-related diseases. In this work, a simple and sensitive electrochemical sandwich-type immunosensing platform was designed by synthesizing b-cyclodextrin (CD) functionalized graphene (CD/GN) hybrid as simultaneously sensing platform and signal transducer coupled with rhodamine b (RhB) as probe. In brief, GN offers large surface area and high conductivity, while CD exhibits superior host-guest recognition capability, thus the primary antibody (Ab1) of HCG can be bound into the cavities of CD/GN to form stable Ab1/CD/GN inclusion complex; meanwhile, the secondary antibody (Ab2) and RhB can also enter into the cavities, producing RhB/Ab2/CD/GN complex. Then, by using Ab1/CD/GN as sensing platform and RhB/Ab2/CD/GN as signal transducer (in which RhB was signal probe), a simple sandwich-type immunosensor was constructed. Under the optimum parameters, the designed immunosensor exhibited a considerable low analytical detection of 1.0 pg mL^-1 and a wide linearity of 0.002 to 10.0 ng mL^-1 for HCG, revealing the developed sandwich-type electrochemical immunosensing platform offered potential real applications for the determination of HCG.

Characterization of the first tuber mustard calmodulin-like gene, BjAAR1, and its functions in responses to abiotic stress and abscisic acid in Arabidopsis

Liuxin Xiang,Yuxian Xia,Ying-Fan Cai,Jijun Liu,Xiaohong He,Quan Sun,Xiaoyan Wang,Yuyin Fu,Yonghong Fan,Daiwen Dong,Guanfan Zhou,Jinjuan Shen,Yihua Liu 한국식물학회 2013 Journal of Plant Biology Vol.56 No.3

The first tuber mustard calmodulin-like (CML) gene BjAAR1 (Brassica juncea var. tumida Tsen et Lee Abiotic stress and Abscisic acid (ABA) Responsive gene 1) was cloned and characterized. The protein encoded by BjAAR1 contains four predicted Ca2+ binding sites (EF-hand motif) and its recombinant protein can bind Ca2+ in vitro. qRT-PCR showed that the expression level of BjAAR1 was rather high in non-swollen stem of tuber mustard and largely reduced in swollen stem. Expression of BjAAR1 enhanced ABA- and stress-induced gene expression in Arabidopsis (Arabidopsis thaliana). Transgenic plants also exhibited hypersensitivity to NaCl, mannitol, and ABA during the seed germination and post-germination stages. ABA biosynthesis inhibitor, norflurazon (NF), rescued hypersensitivity phenotype of transgenic plants to NaCl and mannitol, indicating that BjAAR1 functions in multiple abiotic stresses response through ABA-dependent process.