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.

Sintering of Powder Prepared by Self-Propagating High-Temperature Synthesis (SHS)

Bai Ling,Zhao Xing-Yu,Ge Chang-Chun 한국분말야금학회 2006 한국분말야금학회 학술대회논문집 Vol.2006 No.1

Preparation processing of sintered silicon nitride ceramics was emphatically investigated with Self-Propagating High-Temperature Synthesis (SHS) of silicon nitride prepared by ourselves as raw material. The results indicate that good sinter ability can be obtained with cheaply SHS of silicon nitride preparing silicon nitride materials. The cost of silicon nitride materials will be lowered.

A Quick Query Methods for ONS System based on EPC

Jun Bai,Qiang Zhu,Bai-ling Wang,Liu Yang,Qiang Chen 보안공학연구지원센터 2013 International Journal of Hybrid Information Techno Vol.6 No.6

Down-regulation of FRα Inhibits Proliferation and Promotes Apoptosis of Cervical Cancer Cells in Vitro

Bai, Li-Xia,Ding, Ling,Jiang, Shi-Wen,Kang, Hui-Jie,Gao, Chen-Fei,Chen, Chen,Zhou, Qin,Wang, Jin-Tao Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.14

Folate receptor alpha ($FR{\alpha}$) mediates folate uptake by endocytosis, and while folate is essential to DNA methylation and synthesis and may have an important role in proliferating cells. $FR{\alpha}$ is known to be expressed in rapidly proliferating cells, including many cancer cell lines, but there has been no systematic assessment of expression in cervical cancer cell lines. The aim of the present study was to evaluate the effects of $FR{\alpha}$ on proliferation and apoptosis of cervical cells and correlation mechanism. In this study, we investigated the biological function of $FR{\alpha}$ in Hela cells using RNA interference. Cell proliferation was evaluated by Cell Counting Kit-8 (CCK8) assay, while cell cycling and apoptosis were assessed by flow cytometry, mRNA levels by real time-PCR and protein levels of $FR{\alpha}$, c-Fos and c-Jun by Western blotting. The results revealed that $FR{\alpha}$ was highly expressed in Hela cells and its silencing with a small interfering RNA (siRNA) inhibited cell proliferation and induced cell apoptosis, arresting the cell cycle in G0/G1 stages while decreasing the proportion in S and G2/M stages, and suppressed the expression levels of c-Fos and c-Jun. In conclusion, the results of this study indicated that $FR{\alpha}$ down-regulation might be capable of suppressing cervical cancer cell proliferation and promoting apoptosis. It suggested that $FR{\alpha}$ might be a novel therapeutic target for cervical cancer.

Rethinking Asia: the genealogy of critical thinking in East Asia

Bai Huilin,Pan Hui-Ling Wendy 한국다문화교육학회 2023 Multicultural Education Review Vol.15 No.1

The concept and connotation of critical thinking are formed based on human activities and social development in Europe and America, while relevant studies in East Asia are lacking. How to deconstruct the characteristics, formation and deep reasons of critical thinking in the East based on its continuous and dynamic human activities is still a challenge. Taking China as the research object, we reveal that the East also produces logic-based and intense critical thinking, and gradually integrates the characteristics of inclusiveness, euphemism, and result-oriented noun in the evolution, which can be attributed to the influence of war, religion, and nomadic and agricultural conflicts. This study is of great significance to the in-depth fundamental understanding of oriental critical thinking. It also holds promise to promote critical thinking education in East Asian countries and the cultural exchanges between the East and the West.

Association of a Pre-miR-27a Polymorphism with Cancer Risk: an Updated Meta-analysis

Bai, Rong-Pan,Weng, Yu,Su, Li-Ling,Jin, Ming-Juan,Xu, Zheng-Ping,Lu, Li-Qin,Chen, Guang-Di Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.23

MicroRNA-27a is highly expressed in cancers and has been identified as an oncogenic microRNA. A genetic variant in pre-miR-27a (rs895819) with a transition of A to G has been demonstrated to be associated with cancer risk; however, the results of these studies remain conflicting rather than conclusive. Therefore, we performed a meta-analysis to derive a more precise estimation. Through searching PubMed or other databases up to March 2014 using the following MeSH terms and keywords, "miR-27a", "polymorphism" and "cancer", seventeen case-control studies were identified in this meta-analysis, including 7,813 cases and 9,602. Crude odds ratios (ORs) and corresponding 95% confidence intervals (CIs) were calculated to investigate the association strength between rs895819 and the susceptibility of cancer. The results of the overall meta-analysis did not suggest any association between rs895819 polymorphism and cancer susceptibility, and this remained in Asians as a subgroup. In Caucasians, however, the rs895819 was associated with a reduced cancer risk in heterozygous (OR, 0.83; 95%CI, 0.75-0.93) and dominant models (OR, 0.84; 95%CI, 0.76-0.93), and the [G] allele of rs895819 showed a protective effect (OR, 0.90, 95%CI, 0.84-0.97). Further studies showed a significant association between the [G] allele of rs895819 and decreased risk of breast cancer (0.91; 95%CI, 0.85-0.98), and stratified analyses indicated a protective effect of the [G] allele in Caucasians (OR, 0.89; 95%CI, 0.82-0.98), younger breast cancer cases (OR, 0.87; 95%CI, 0.79-0.96), and in the group of unilateral breast cancer patients (OR, 0.90; 95%CI, 0.83-0.97). These findings suggest an association between pre-miR-27a polymorphism rs895819 and cancer risk in Caucasians. The protective effect of rs895819 [G] allele in younger breast cancer and in the group of unilateral breast cancer patients await further confirmation since the included studies in this meta-analysis were limited.

Folate Deficiency and FHIT Hypermethylation and HPV 16 Infection Promote Cervical Cancerization

Bai, Li-Xia,Wang, Jin-Tao,Ding, Ling,Jiang, Shi-Wen,Kang, Hui-Jie,Gao, Chen-Fei,Chen, Xiao,Chen, Chen,Zhou, Qin Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.21

Fragile histidine triad (FHIT) is a suppressor gene related to cervical cancer through CpG island hypermethylation. Folate is a water-soluble B-vitamin and an important cofactor in one-carbon metabolism. It may play an essential role in cervical lesions through effects on DNA methylation. The purpose of this study was to observe effects of folate and FHIT methylation and HPV 16 on cervical cancer progression. In this study, DNA methylation of FHIT, serum folate level and HPV16 status were measured using methylation-specific polymerase chain reaction (MSP), radioimmunoassay (RIA) and polymerase chain reaction (PCR), respectively, in 310 women with a diagnosis of normal cervix (NC, n=109), cervical intraepithelial neoplasia (CIN, n=101) and squamous cell carcinoma of the cervix (SCC, n=101). There were significant differences in HPV16 status (${\chi}^2=36.64$, P<0.001), CpG island methylation of FHIT (${\chi}^2=71.31$, P<0.001) and serum folate level (F=4.57, P=0.011) across the cervical histologic groups. Interaction analysis showed that the ORs only with FHIT methylation (OR=11.47) or only with HPV 16 positive (OR=4.63) or with serum folate level lower than 3.19ng/ml (OR=1.68) in SCC group were all higher than the control status of HPV 16 negative and FHIT unmethylation and serum folate level more than 3.19ng/ml (OR=1). The ORs only with HPV 16 positive (OR=2.58) or with serum folate level lower than 3.19ng/ml (OR=1.28) in CIN group were all higher than the control status, but the OR only with FHIT methylation (OR=0.53) in CIN group was lower than the control status. HPV 16 positivity was associated with a 7.60-fold increased risk of SCC with folate deficiency and with a 1.84-fold increased risk of CIN. The patients with FHIT methylation and folate deficiency or with FHIT methylation and HPV 16 positive were SCC or CIN, and the patients with HPV 16 positive and FHIT methylation and folate deficiency were all SCC. In conclusion, HPV 16 infection, FHIT methylation and folate deficiency might promote cervical cancer progression. This suggests that FHIT may be an effective target for prevention and treatment of cervical cancer.

tae-miR9674a, a microRNA member of wheat, confers plant drought and salt tolerance through modulating the stomata movement and ROS homeostasis

Wang Ling,Bai Xinyang,Qiao Yuanjinzi,Si Lili,Yu Zidi,Ni Chenyang,Li Tianjiao,Guo Chengjin,Xiao Kai 한국식물생명공학회 2023 Plant biotechnology reports Vol.17 No.4

The members of the microRNA (miRNA) family exert essential roles in modulating plant growth and development as well as responses to diverse stresses, through negatively regulating their target genes at posttranscriptional or translational levels. In this study, we characterized taemiR9674a, a miRNA member in T. aestivum, in mediating plant responses to drought and salt stresses. Seven genes in total were predicted to act as the targets of tae-miR9674a via modulation of transcript cleavage. The transcripts of tae-miR9674a in roots and leaves were response to both stresses of drought and salt, displaying to be gradually upregulated following the progression of a 27-h regime of above stress treatments. The transgenic tobacco lines of tae-miR9674a exhibited modified growth traits under drought and salt treatments. Of these, the line with miRNA overexpression (i.e., Sen 1) improved drastically on plant biomass, leaf area, and root length, whereas that with its knockdown expression (Anti 1) significantly alleviated on above growth traits compared with wild type. The modified stress responses of tae-miR9674a were shown to be closely associated with the role of miRNA in regulating a suite of physiological parameters, of which Sen 1 displayed improved osmotic stress defensive-related traits, such as fastened stomata closing rate, increased leaf water retention capacity, enhanced osmolytes contents, and elevated antioxidant enzyme (AE) activities. The expression of NtP5CS1 involving proline biosynthesis and NtFeSOD, NtCAT1 and NtPOD4, the AE genes involved in modulating ROS homeostasis, was upregulated in Sen 1 upon drought and salt stresses, suggesting their involvement in miRNA-mediated plant drought and salt responses. Transcriptome analysis indicated that tae-miR9674a leads to modified expression of quantities of genes that functionally associate with GO terms “biological process”, “cellular component”, and “molecular function”, which are overrepresented by the biochemical pathways of phytohormones (i.e., ethylene and jasmonic acid), salt response, salt/drought osmotic stress response through abscisic acid-dependent pathway and reactive oxygen species (ROS) homeostasis. Our investigation suggested that tae-miR9674a is an essential mediator in plant osmotic stress tolerance by positively regulating osmotic stress acclimation, cellular ROS homeostasis, and related defensive processes.

Knock-down of OsLOX by RNA interference leads to improved seed viability in rice

Suyang Bai,Niqing He,Lu Zhou,Beibei Shen,Wei Wu,Xi Liu,Ling Jiang,Jianmin Wan 한국식물학회 2015 Journal of Plant Biology Vol.58 No.5

Previous work found that lipoxygenases were key enzymes in lipid peroxidation, which causes grain deterioration during storage. In order to obtain better seed viability in rice, 10 marker-free knock-down lines were obtained in the progeny of endogenous OsLOX knock-down mutations caused by the RNAi technology. After artificial accelerated aging, there were four types of knock-down lines with higher seed viability than wild type (receptor parent). OsLOX3 knock-down line NPF1 was of special interest. In a series of experiments, including Southern blots, analysis of OsLOX3 expression, and enzymatic activity, NPF1 had better seed viability than wild-type. We also investigated the main agronomic characters of both knock-down lines and non-transgenetic wild type families. Knock-down lines were identified with generally excellent agronomic characteristics similar to the wild-type.

Transient simulation and experiment validation on the opening and closing process of a ball valve

Yong Han,Ling Zhou,Ling Bai,Peng Xue,Wanning Lv,Weidong Shi,Gaoyang Huang 한국원자력학회 2022 Nuclear Engineering and Technology Vol.54 No.5

The ball valve is an important device in the pipeline transportation system of nuclear power plants. Itsoperational stability and safety directly affect the normal working of nuclear power plants. In this study,the transient numerical simulation of the opening and closing process of a ball valve was conducted onthe basis of the flow interruption capability experiment of the ball valve by using the moving meshmethod and inlet and outlet variable boundary conditions. The flow rate and pressure difference withtime of the opening and closing process of the ball valve were studied. The internal flow characteristics ofthe ball valve under different relative openings were analyzed in conjunction with the typical back-stepflow structure. Results show that the transient numerical results agree well with the experimental results. The internal flow characteristics of the ball valve are similar at the same opening during openingand closing process. At small opening, the spool and outlet channels easily form a back-step flowstructure. The disappearance and generation of backflow vortices during opening and closing occur at85% opening and 75% opening, respectively. With the decrease in opening degree, the difference invortex core area in the flow channel of the ball valve spool in the opening and closing process graduallyappears. The research results provide some reference value for the design and optimization of ball valves.