Selection and evaluation of reference genes for gene expression using quantitative real‐time PCR in Mythimna separata walker (Lepidoptera: Noctuidae)

Bai‐Zhong Zhang,Jun-Jie LIU,Xi-Ling CHEN,Guo-Hui YUAN 한국곤충학회 2018 Entomological Research Vol.48 No.5

In order to precisely assess gene expression levels, the suitable internal reference genes must be served to quantify real‐time reverse transcription polymerase chain reaction (RT‐qPCR) data. For armyworm, Mythimna separata, which reference genes are suitable for assessing the level of transcriptional expression of target genes have yet to be explored. In this study, eight common reference genes, including β‐actin (β‐ACT), 18 s ribosomal (18S), 28S ribosomal (28S), glyceraldehyde‐3‐phosphate (GAPDH), elongation fator‐alpha (EF1α), TATA box binding protein (TBP), ribosomal protein L7 (RPL7), and alpha‐tubulin (α‐TUB) that in different developmental stages, tissues and insecticide treatments of M. separata were evaluated. To further explore whether these genes were suitable to serve as endogenous controls, three software‐based approaches (geNorm, BestKeeper, and NormFinder), the delta Ct method, and one web‐based comprehensive tool (RefFinder) were employed to analyze and rank the tested genes. The optimal number of reference genes was determined using the geNorm program, and the suitability of particular reference genes was empirically validated according to normalized HSP70, and MsepCYP321A10 gene expression data. We found that the most suitable reference genes for the different experimental conditions. For developmental stages, 28S/RPL7 were the optimal reference genes, both RPL7/EF1α were suitable for experiments of different tissues, whereas for insecticide treatments, 28S/α‐TUB were suitable for normalizations of expression data. In addition, 28S/α‐TUB were the suitable reference genes because they have the most stable expression among different developmental stages, tissues and insecticide treatments. Our work is the first report on reference gene selection in M. separata, and might serve as a precedent for future gene expression studies.

Cough Inhibition Activity of Schisandra chinensis in Guinea Pigs

Shan Zhong,Li-Ping Bai,Xiao-Dong Liu,Die-Yi Cai,Lee-Fong Yau,Chu-Qin Huang,Jia-Qi Zhang,Ke-Fang Lai,Nan-Shan Zhong 한국식품영양과학회 2021 Journal of medicinal food Vol.24 No.4

Chronic cough is very common in respiratory clinics, and no effective drugs are available. Schisandra chinensis (Turcz.) Baill. (S. chinensis), an important traditional Chinese medicine, has been extensively prescribed for patients with a persistent cough. Preliminary research indicated that 95% ethanol extracts (EE) of S. chinensis showed remarkable antitussive activity in guinea pigs exposed to cigarette smoke (CS). To find out the antitussive ingredients of S. chinensis, EE was divided into four fractions according to the polarity: petroleum ether extract (PEE), ethyl acetate extract (ECE), n-butyl alcohol extract, and residue extract. The antitussive, antioxidant, and anti-inflammatory effects of the four fractions were evaluated in a guinea pig model of cough hypersensitivity induced by CS exposure. Eighteen main constituents of the two effective fractions, PEE and ECE, were identified using ultra-high-pressure liquid chromatography electronic spray ion time-of-flight mass spectrometry. The cough inhibition activities of compound 1, 3, 9, 10, 17 were evaluated on citric acid induced acute cough guinea pigs. The results showed that the antitussive activity of EE was almost all contained in PEE and ECE. The 16 major peaks in PEE were identified as 15 lignans (1–12 and 14–16) and 1 triterpene (compound 13), and 3 major peaks (1, 17, and 18) in ECE were also identified as lignans. Three doses of five compounds brought about a significant decrease in number of cough efforts (P < .01), and the cough inhibition rates were between 40.9% and 85.1%. Therefore, lignans are the antitussive ingredients of S. chinensis.

Promoter demethylation mediates the expression of ZNF645, a novel cancer/testis gene

( Gang Bai ),( Yun Qiang Liu ),( Hao Zhang ),( Dan Su ),( Da Chang Tao ),( Yuan Yang ),( Yong Xin Ma ),( Si Zhong Zhang ) 생화학분자생물학회 (구 한국생화학분자생물학회) 2010 BMB Reports Vol.43 No.6

Cancer/testis (CT) antigens exhibit highly tissue-restricted expression and are considered promising targets for cancer vaccines. Here we identified a novel CT gene ZNF645 which restrictively expresses in normal human testes and lung cancer patients (68.3%). To investigate the promoter methylation status of ZNF645, we carried out bisulfite genomic sequencing and found that the CpG island in its promoter was heavily methylated in normal lung tissues without the expression of ZNF645, whereas there was high demethylation in normal human testes and lung carcinoma tissues with its expression. Also ZNF645 could be remarkably activated in A549 and HEK293T cells treated by DNA demethylation agent 5`-aza-2`-deoxycytidine. And the dual luciferase assay revealed that the promoter activity of the ZNF645 was inhibited by methylation of the CpG island region. Therefore, we proposed that ZNF645 is a CT gene and activated in human testis and lung cancers by demethylation of its promoter region. [BMB reports 2010; 43(6): 400-406]

Dietary maifanite supplementation did not affect the apparent total tract digestibility of calcium and phosphorus in growing pigs

Li Li Bai,Dong Xu Ming,Shu Ren Dong,Zhong Yue Yang,Wen Hui Wang,Shuai Zhang,Xiang Shu Piao,Ling Liu,Fenglai Wang 아세아·태평양축산학회 2018 Animal Bioscience Vol.31 No.2

Objective: This study was conducted to determine the effects of dietary maifanite supplementation and fecal collection method on the apparent total tract digestibility (ATTD) of calcium (Ca) and phosphorus (P) and blood parameters in growing pigs. Methods: Thirty-six growing barrows (Duroc×Landrace×Yorkshire; 27.0±2.6 kg) were allotted to six dietary treatments with 6 pigs per treatment according to body weight in a completely randomized design. The experimental treatments were: i) Low Ca+cornstarch (2.25%), ii) Low Ca+maifanite (2.25%), iii) Medium Ca+cornstarch (1.42%), iv) Medium Ca+maifanite (1.42%), v) High Ca+cornstarch (0.64%), and vi) High Ca+maifanite (0.64%). Feces were collected by the total collection (TC) and indicator method (IM). At the beginning and the end of the experiment, blood samples were collected from each pig. Results: For the TC method, there were no difference in Ca intake, fecal Ca output, Ca retention and the ATTD of Ca between cornstarch and maifanite diets at the same dietary Ca level. However, urinary Ca excretion was lower (p = 0.01) in pigs fed low Ca diets without maifanite supplementation compared with other dietary treatments. Dietary maifanite supplementation had no effect on the P metabolism in growing pigs. For the IM method, there was no difference in Ca digestibility between cornstarch and maifanite diets at the same dietary Ca level. The ATTD of P was greater (p<0.01) in pigs fed the high Ca diet with maifanite supplementation compared with the high Ca diet with cornstarch treatment. Dietary inclusion of maifanite had no effect on blood parameters in growing pigs. Conclusion: Dietary maifanite supplementation had no effect on the ATTD of Ca and P and serum parameters in growing pigs. The IM resulted in lower digestibility values than the TC method.

A Novel Self-Organized Fuzzy Neural Network Surface Reconstruction Algorithm for Point Clouds Without Normal

Liu Yan-ju,Liu Yan-zhong,Tao Bai-rui,Jiang Jin-gang,Zhang Hong-lie 보안공학연구지원센터 2014 International Journal of Database Theory and Appli Vol.7 No.4

Reduction Strategy of Point Clouds to Reconstruct Surface Based on Fuzzy Clustering

Liu Yan-ju,Jiang Jin-gang,Tao Bai-rui,Zhang Hong-lie,Liu Yan-zhong 보안공학연구지원센터(IJGDC) 2014 International Journal of Grid and Distributed Comp Vol.7 No.4

De novo assembly of the transcriptome for Greenbug (Schizaphis graminum Rondani) and analysis on insecticide resistance-related genes

Jun-Jie LIU,Liu-Yang LU,Hui-Hui LIU,Ya-She LI,Xu SU,Bai-Zhong ZHANG,Xi-Ling CHEN 한국곤충학회 2019 Entomological Research Vol.49 No.8

The greenbug, Schizaphis graminum Rondani is a major pest species of wheat crops. In this study, a transcriptome sequencing, and the expression of the 12 genes related to insecticide resistance were conducted in S. graminum. The sequencing and subsequent bioinformatics analysis outputed 46,593 unigenes, among which 28,289 unigenes were annotated to corresponding functions by blasting with high homologous genes in database, giving annotation rate of 60.72%. To gain insight into the mechanism of insecticide resistance, the expression of the 12 genes related to insecticide resistance for S. graminum was investigated. The expression level of aminopeptidase N (AN), cytochrome P450 (CYP), acetylcholinesterase 1 (AC), catalase (CAT), cytochrome c oxidas (CCC), GABA receptor (GABA), glutathione S-transferase (GST) were highest in the apterous nymphs among different developmental stages; The expression level of AN, CBL, CYP, CA, SD, and GST were relatively more abundant in the abdomen compared to head and throax. The results could give out the key information about the relationship between the expression of these genes in different developmental stages, tissues, treatments and metabolism of insecticides. These genes that were co-up-regulated significantly in third instar nymphs of S. graminum induced by imidacloprid, were consistent with their putative involvement in insecticide resistance. This provides most comprehensive transcriptome data for S. graminum to further study and managenment.