Correlation between DNA methylation and Thymic Stromal Lymphopoietin expression in asthmatic airway epithelial cells

Yan‑Li Li,Xi‑Qian Xing,Yi Xiao,Yan‑Hong Liu,Yu‑Shan Zhou,Min Zhuang,Chao‑Qian Li 한국유전학회 2020 Genes & Genomics Vol.42 No.12

Background: The overexpression of TSLP and DNA methylation in asthma were both risk factors the relationship was not clear. Objective: This study aimed to investigate the relationship between methylation status of TSLP promoter and mRNA/protein expression in asthmatic airway epithelial cells. Methods: Human bronchial epithelial cells were cultured in vitro and divided into: Control group, treated with PBS, model group, sensitized with LPS (10 μg/mL) for 12 h (37 °C, 5% CO2). Other groups were cultured with the pCMV3 plasmid (M + NC/pCMV), pGPH1 plasmid (M + NC/pGPH), DNMT1/pCMV3 plasmid (M + DNMT1/pCMV), and DNMT1/pGPH1 plasmid (M + DNMT1/pGPH) for 48 h. The expression of DNA methyltransferase 1 and TSLP were measured using real-time PCR and western blotting. Results: Compared with the control group, TSLP mRNA (1.00 ± 0.00 vs. 2.82 ± 0.81 vs. 1, P < 0.001) and protein (1.07 ± 0.04 vs. 1.46 ± 0.11, P < 0.01) were significantly greater, and the methylation of promoter was lower (92.75 ± 1.26 vs. 58.57 ± 3.34, P < 0.05) in the model group. Compared with the model group, TSLP mRNA (2.82 ± 0.81 vs. 1.17 ± 0.10, P < 0.001) decreased, but TSLP promoter methylation increased (58.57 ± 3.34 vs. 92.58 ± 7.30, P < 0.05) in M + DNMT1/pCMV. TSLP mRNA and protein were higher (2.82 ± 0.81 vs. 5.32 ± 0.21, P < 0.001; 1.46 ± 0.11 vs. 1.94 ± 0.11, respectively, P < 0.01), TSLP promoter methylation was lower (58.57 ± 3.34 vs. 33.57 ± 4.29, P < 0.05) in M + DNMT1/pGPH. Conclusions: Overexpression of TSLP in asthmatic airway epithelial cells may be regulated by DNA demethylation.

Characterization of the Methylation Status of Pax7 and Myogenic Regulator Factors in Cell Myogenic Differentiation

Chao, Zhe,Zheng, Xin-Li,Sun, Rui-Ping,Liu, Hai-Long,Huang, Li-Li,Cao, Zong-Xi,Deng, Chang-Yan,Wang, Feng Asian Australasian Association of Animal Productio 2016 Animal Bioscience Vol.29 No.7

Epigenetic processes in the development of skeletal muscle have been appreciated for over a decade. DNA methylation is a major epigenetic modification important for regulating gene expression and suppressing spurious transcription. Up to now, the importance of epigenetic marks in the regulation of Pax7 and myogenic regulatory factors (MRFs) expression is far less explored. In the present study, semi-quantitative the real-time polymerase chain reaction (RT-PCR) analyses showed MyoD and Myf5 were expressed in activated and quiescent C2C12 cells. MyoG was expressed in a later stage of myogenesis. Pax7 was weakly expressed in differentiated C2C12 cells. To further understand the regulation of expression of these genes, the DNA methylation status of Pax7, MyoD, and Myf5 was determined by bisulfite sequencing PCR. During the C2C12 myoblasts fusion process, the changes of promoter and exon 1 methylation of Pax7, MyoD, and Myf5 genes were observed. In addition, an inverse relationship of low methylation and high expression was found. These results suggest that DNA methylation may be an important mechanism regulating Pax7 and MRFs transcription in cell myogenic differentiation.

A Fiber Model Based on Secondary Development of ABAQUS for Elastic–Plastic Analysis

Yan-Li Shi,Hua-Wei Li,Wen-Da Wang,Chao Hou 한국강구조학회 2018 International Journal of Steel Structures Vol.18 No.5

With the aim to provide an effi cient platform for the elastic–plastic analysis of steel structures, reinforced concrete (RC) structures and steel–concrete composite structures, a program iFiberLUT based on the fi ber model was developed within the framework of ABAQUS. This program contains an ABAQUS Fiber Generator which can automatically divide the beam and column cross sections into fi ber sections, and a material library which includes several concrete and steel uniaxial material models. The range of applications of iFiberLUT is introduced and its feasibility is verifi ed through previously reported test data of individual structural members as well as planar steel frames, RC frames and composite frames subjected to various loadings. The simulation results indicate that the developed program is able to achieve high calculation accuracy and favorable convergence within a wide range of applications.

Risk Factors for Surgical Site Infections after Liver Resec-tion (A Multivariate Analysis of 6,132 Patients)

( Li-yang Sun ),( Jiong-jie Yu ),( Ju-dong Li ),( Xin-fei Xu ),( Jia-he Wang ),( Bing Quan ),( Wen-tao Yan ),( Feng Shen ),( Chao Li ),( Lei Liang ),( Tian Yang ) 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: Surgical site infection (SSI) is a common postoperative complication and associated with an increased morbidity, hospital stay, and overall cost. The aim of the present study was to identify risk factors for SSIs after hepatic resection based on a large single-center cohort. Methods: A retrospective study was conducted of 6,132 patients who underwent liver resection without concomitant biliary reconstruction or gastrointestinal procedures between 2014 and 2016 at the largest hepatic center in China. The occurrences of SSI, classified as incisional SSI and organ/space SSI within 30 days after operation were investigated. Patient- and surgical-related risk variables were collected using standardized data collection form. A likelihood ratio forward regression model was used to assess the independent association of risk factors with SSI. Results: SSI developed in 587 patients (9.6%), including superficial/deep incisional SSI in 357 patients (5.8 %), and organ/ space SSI in 304 patients (5.0 %). Multivariate logistic regression analysis showed that obesity, diabetes mellitus, ASA score ≥ 2, liver cirrhosis, re-hepatectomy, hepatoliathiasis, and intraoperative blood transfusion were independent risk factors of overall SSI. However, incisional and organ/space SSI differed from each other with respect to risk factors. Among a variety of risk factors, hepatolithiasis, liver cirrhosis, and intraoperative blood transfusion were consistently associated with both incisional and organ/space SSI. Conclusions: SSI is a common complication after liver resection, and more caution should be taken in patients with hepatolithiasis or liver cirrhosis. Prevention strategies focusing on factors associated with SSI is necessary in order to reduce SSI after liver resection.

IOACHIMESCU'S CONSTANT

CHAO-PING CHEN,Li,Xu,LI LI,YAN-QIN XU 장전수학회 2010 Proceedings of the Jangjeon mathematical society Vol.13 No.3

The Ioachimescu's constant I = 0.539645.... is defined as the limit of the sequence In = [수식] (n ∈ N). In this paper, we establish the asymptotic representation of the sequence (In)n∈N, and obtain the upper and lower bounds for In- and I.

Selection of Reference Genes for Real-time Quantitative PCR Normalization in the Process of Gaeumannomyces graminis var. tritici Infecting Wheat

Li-hua Xie,Xin Quan,Jie Zhang,Yan-yan Yang,Run-hong Sun,Ming-cong Xia,Bao-guo Xue,Chao Wu,Xiao-yun Han,Ya-nan Xue,Li-rong Yang 한국식물병리학회 2019 Plant Pathology Journal Vol.35 No.1

Gaeumannomyces graminis var. tritici is a soil borne pathogenic fungus associated with wheat roots. The accurate quantification of gene expression during the process of infection might be helpful to understand the pathogenic molecular mechanism. However, this method requires suitable reference genes for transcript normalization. In this study, nine candidate reference genes were chosen, and the specificity of the primers were investigated by melting curves of PCR products. The expression stability of these nine candidates was determined with three programs-geNorm, Norm Finder, and Best Keeper. TUBβ was identified as the most stable reference gene. Furthermore, the exopolygalacturonase gene (ExoPG) was selected to verify the reliability of TUBβ expression. The expression profile of ExoPG assessed using TUBβ agreed with the results of digital gene expression analysis by RNA-Seq. This study is the first systematic exploration of the optimal reference genes in the infection process of Gaeumannomyces graminis var. tritici.

Evaluation of reference genes for miRNA expression analysis in Galeruca daurica (Coleoptera: Chrysomelidae) using qRT‐PCR

Hai-Chao WANG,Ling LI,Yan-Yan LI,Yao TAN,Bao-Ping PANG 한국곤충학회 2021 Entomological Research Vol.51 No.8

The selection of suitable reference genes (RGs) for data normalization is a crucial step for obtaining reliable results from gene expression analysis using the quantitative real-time polymerase chain reaction (qRT-PCR). Expression stability of eleven putative RGs (miR-100-5p, miR-92a-3p, miR-998-3p, miR-279d-3p, miR-305-5p, miR-276a-3p, miR-275-3p, miR-9a-5p, miR-2a-3p, U6 snRNA and 5S rRNA) was assessed by using a web-based tool: RefFinder, which integrates four algorithms (Delta Ct, geNorm, NormFinder, and BestKeeper), and the best combination number of RGs was calculated by geNorm. Validation of selected RGs was performed by an expression analysis of let-7-5p under different experimental conditions. The best combinations of RGs were shown as follows: miR-100-5p + miR-275-3p for temperature treatment, miR-100-5p + miR-305-5p for adult tissues, miR- 276a-3p + miR-9a-5p for genders, miR-2a-3p + miR-276a-3p for days after eclosion, U6 snRNA + miR-9a-5p + miR-2a-3p for developmental stages, and U6 snRNA + miR-279d-3p for all above conditions, and there was no one RG suitable for all experimental conditions. Incorrect RG selection affected the evaluation of let-7-5p abundance. Our findings demonstrate that an integrated approach of multiple algorithms is necessary to identify reliable, stable RGs for miRNA evaluation, and provide a basis for further studies on expression and function analysis of miRNAs and their target genes in Galeruca daurica, a new pest with great outbreaks in Inner Mongolia since 2009.

Pretreatment Serum Amyloid A and C-reactive Protein Comparing with Epstein-Barr Virus DNA as Prognostic Indicators in Patients with Nasopharyngeal Carcinoma: A Prospective Study

Qiu-Yan Chen,Qing-Nan Tang,Lin-Quan Tang,Wen-Hui Chen,Shan-Shan Guo,Li-Ting Liu,Chao-Feng Li,Yang Li,Yu-Jing Liang,Xue-Song Sun,Ling Guo,Hao-Yuan Mo,Rui Sun,Dong-Hua Luo,Yu-Ying Fan,Yan He,Ming-Yuan C 대한암학회 2018 Cancer Research and Treatment Vol.50 No.3

Purpose The measuring Epstein-Barr virus (EBV) DNA is an important predictor of nasopharyngeal carcinoma (NPC). This study evaluated the predictive value of pretreatment serum amyloid A (SAA) and C-reactive protein (CRP) comparing with EBV DNA in patients with NPC. Materials and Methods In an observational study of 419 non-metastatic NPC patients, we prospectively evaluated the prognostic effects of pretreatment SAA, CRP, and EBV DNA on survival. The primary endpoint was progress-free survival (PFS). Results The median level of SAA and CRP was 4.28 mg/L and 1.88 mg/L, respectively. For the high- SAA group (> 4.28 mg/L) versus the low-SAA ( 4.28 mg/L) group and the high-CRP group (> 1.88 mg/L) versus the low-CRP ( 1.88 mg/L) group, the 5-year PFS was 64.5% versus 73.1% (p=0.013) and 65.2% versus 73.3% (p=0.064), respectively. EBV DNA detection showed a superior predictive result, the 5-year PFS in the EBV DNA  1,500 copies/mL group was obviously different than the EBV DNA < 1,500 copies/mL group (62.2% versus 77.8%, p < 0.001). Multifactorial Cox regression analysis confirmed that in the PFS, the independent prognostic factors were including EBV DNA (hazard ratio [HR], 1.788; p=0.009), tumour stage (HR, 1.903; p=0.021), and node stage (HR, 1.498; p=0.049), but the SAA and CRP were not included in the independent prognostic factors. Conclusion The results of SAA and CRP had a certain relationship with the prognosis of NPC, and the prognosis of patients with high level of SAA and CRP were poor. However, the predictive ability of SAA and CRP was lower than that of EBV DNA.

Selection of Reference Genes for Real-time Quantitative PCR Normalization in the Process of Gaeumannomyces graminis var. tritici Infecting Wheat

Xie, Li-hua,Quan, Xin,Zhang, Jie,Yang, Yan-yan,Sun, Run-hong,Xia, Ming-cong,Xue, Bao-guo,Wu, Chao,Han, Xiao-yun,Xue, Ya-nan,Yang, Li-rong The Korean Society of Plant Pathology 2019 Plant Pathology Journal Vol.35 No.1

Gaeumannomyces graminis var. tritici is a soil borne pathogenic fungus associated with wheat roots. The accurate quantification of gene expression during the process of infection might be helpful to understand the pathogenic molecular mechanism. However, this method requires suitable reference genes for transcript normalization. In this study, nine candidate reference genes were chosen, and the specificity of the primers were investigated by melting curves of PCR products. The expression stability of these nine candidates was determined with three programs-geNorm, Norm Finder, and Best Keeper. $TUB{\beta}$ was identified as the most stable reference gene. Furthermore, the exopolygalacturonase gene (ExoPG) was selected to verify the reliability of $TUB{\beta}$ expression. The expression profile of ExoPG assessed using $TUB{\beta}$ agreed with the results of digital gene expression analysis by RNA-Seq. This study is the first systematic exploration of the optimal reference genes in the infection process of Gaeumannomyces graminis var. tritici.

Combined Expression of Metastasis Related Markers Naa10p, SNCG and PRL-3 and its Prognostic Value in Breast Cancer Patients

Min, Li,Ma, Ruo-Lan,Yuan, Hua,Liu, Cai-Yun,Dong, Bing,Zhang, Cheng,Zeng, Yan,Wang, Li,Guo, Jian-Ping,Qu, Li-Ke,Shou, Cheng-Chao Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.7

Combinations of multiple biomarkers representing distinct aspects of metastasis may have better prognostic value for breast cancer patients, especially those in late stages. In this study, we evaluated the protein levels of N-${\alpha}$-acetyltransferase 10 protein (Naa10p), synuclein-${\gamma}$ (SNCG), and phosphatase of regenerating liver-3 (PRL-3) in 365 patients with breast cancer by immunohistochemistry. Distinct prognostic subgroups of breast cancer were identified by combination of the three biomarkers. The Naa10p+SNCG-PRL-3-subgroup showed best prognosis with a median distant metastasis-free survival (DMFS) of 140 months, while the Naa10p-SNCG+PRL-3+subgroup had the worst prognosis with a median DMFS of 60.5 months. Multivariate analysis indicated Naa10p, SNCG, PRL-3, and the TNM classification were all independent prognostic factors for both DMFS and overall survival (OS). The three biomarker combination of Naa10p, SNCG and PRL-3 performed better in patients with lymph node metastasis, especially those with more advanced tumors than other subgroups. In conclusion, the combined expression profile of Naa10p, SNCG and PRL-3, alone or in combination with the TNM classification system, may provide a precise estimate of prognosis of breast cancer patients.