A Multiplex Sensitive Quantification of MicroRNAs Based on Competitive PCR

Maochun Wang,Li Tong,Sijia Wang,Kai Li,Junhua Xiao,Yuxun Zhou 한국생물공학회 2017 Biotechnology and Bioprocess Engineering Vol.22 No.1

MicroRNAs (miRNAs) are endogenous ~22 nt RNAs that play important regulatory roles in animals and plants by targeting mRNAs for cleavage or gene silencing. Although quantitative real-time PCR (qRT-PCR) had been widely used for miRNAs quantification, a multiplex quantification method is demanding. In this study, we successfully detected 2 miRNAs (miR-505-3p and miR- 21a-5p) and an internal control (miR-16-5p) with only one reaction based on competitive PCR (cPCR) with high sensitivity. For each miRNA, two stem-loop reverse transcription (RT) primers were designed to produce two different templates: the competitor cDNA and the target cDNA, which had similar sequences except for 3 nucleotides different in length. RNA from a control sample was reverse transcribed with the competitive RT primers of multiple genes. Samples for test were reverse transcribed with target RT primers to obtain target cDNAs. Target cDNA was mixed with competitor cDNA to be used as the template for a multiplex fluorescent cPCR reaction. The cPCR products were separated on polyacrylamide gel electrophoresis with ABI 377 DNA sequencer and each fluorescent peak was quantified by its intensity. In this method, we compared the expression level of miR-505-3p in two tissues (thalamus and tail) between C57BL/6J and C3H/HeJ mice. The results showed that in the thalamus, which had high abundance of miR-505-3p, both cPCR and SYBR Green based qRT-PCR provided a sensitive quantification outcome. However, in the tail, which had extremely low level of miR-505-3p, it could be steadily detected by cPCR even after 8 times dilution with a relatively high sensitivity, while qRT-PCR can’t detect any product only after 2 times dilution. The variation as low as 12.2% between samples could be clarified by cPCR, which could not be accomplished by qRT-PCR. This method enables multiplex, accurate and sensitive quantification of miRNAs with fewer precious RNA samples than qRT-PCR.

드라마 OST 출시 영향요인

왕사가(Wang, SiJia),신형덕(Shin, Hyung-Deok) 한국문화산업학회 2020 문화산업연구 Vol.20 No.4

배경음악(Original Sound Track, 이하 OST)은 영화, 드라마, 애니메이션, 게임 등 문화콘텐츠에 사용되어 부가가치를 창출한다. 특히 TV 드라마에 사용되는 OST는 등장인물별과 테마별로 세분화되어 만들어지기도 하고 등장인물이 직접 부르기도 하는 등 드라마의 경쟁력을 높이는 중요한 수단으로 활용되고 있다. 그러나 효과적인 OST 활용의 하나의 조건인 OST의 출시 조건에 대한 연구는 아직 이루어지지 않았다. 본 연구는 2017년 한국에서 방영된 18개 드라마의 294개 회차를 대상으로 드라마 OST 출시에 영향에 미치는 요인에 대해 조사하였다. 그 결과 드라마가 중반부에 속한 경우와 드라마 주연배우의 스타파워가 높을 때에 OST 출시 경향이 높다는 것을 발견하였다. 그러나 드라마 방영 직전회차 시청률은 OST 출시에 영향을 미치지 않았다. 본 연구는 드라마 제작자는 OST의 출시 시점을 통해 드라마의 경쟁력을 관리할 수 있다는 실무적 교훈을 발견함과 동시에 시청률과 OST 출시와의 관계는 존재하지 않는다는 점을 확인했다. 이러한 결과는 드라마 제작자에게 드라마의 경쟁력에 대한 실무적 시사점을 던짐과 동시에 OST 관련 분야 실증분석의 하나의 예를 보여준다. Original Sound Track(OST) creates value when used in cultural contents such as movies, dramas, animations, and games. Especially, OSTs can enhance competitive advantages of TV dramas when they are customized for various characters and themes, and when major characters sing the OST themselves. However, research on OST release conditions for effective OST utilization has not been made. Using 294 drama broadcasting data among 18 dramas released in Korea in 2017, this study investigated relationship between drama elements such as storytelling steps, star power and audience rating, and the event of OST release. This study found that OST is likely to be released when the story of the drama is in the middle stages and the drama has star power. In addition, audience rating of the recent episode of the drama did not affect OST release.

인공 지능을 이용한 부산해양박물관 체험을 통한 발전방안 연구

왕스가(SiJia Wang),김면(Myun Kim) 한국디자인학회 2024 한국디자인학회 학술발표대회 논문집 Vol.2024 No.6

Synthesis and Properties of Benzoxazine Monomers Bearing Both 3-Methyltetrahydrophtalimide and Nitrile Groups: Para-Para vs. Ortho-Ortho

Yuting Wang,Sijia You,Jie Hu,Kan Zhang 한국고분자학회 2020 Macromolecular Research Vol.28 No.1

Two 3-methytetrahydrophtalimide-functional benzoxazine monomers containing nitrile group have been prepared in order to achieve the basic design rules for developing high-performance thermosets via multifunctional benzoxazines. The chemical structures of the two new synthesized benzoxazine monomers have been confirmed by 1H NMR, 13C NMR and FT-IR. Besides, the polymerization behavior has been studied by using DSC and in situ FT-IR. In addition, the thermal stability and flame retardant properties of the resulting polybenzoxazines have also been investigated using TGA and MCC. The polybenzoxazine based on ortho-3-methytetrahydrophtalimide functional benzoxazine containing ortho-nitrile group exhibits excellent thermal stability and low flammability, with a Td10 of 378 °C and a HRC value of 180 J/(g K).

New Construction of Efficient Hierarchical Identity Based Signature in the Standard Model

He Liu,Sijia Wang,Mangui Liang 보안공학연구지원센터 2013 International Journal of Security and Its Applicat Vol.7 No.4

New Construction of Efficient Certificateless Aggregate Signatures

He Liu,Sijia Wang,Mangui Liang,Yongqian Chen 보안공학연구지원센터 2014 International Journal of Security and Its Applicat Vol.8 No.1

Effect of Shadow Banking Activities on Firm Risk and Performance: Entrusted Loan Evidence from Chinese Listed Firms

Meng-Wen Wu,Xiaoyan Wang,Sijia Qiao,Chung-Hua Shen,Juan Wang 한국증권학회 2022 Asia-Pacific Journal of Financial Studies Vol.51 No.2

This study investigates the effect of non-financial firms’ activities in shadow banking on firm risk and performance. Using manually collected data of entrusted loans from Chinese listed firms, we find that lending firms’ bankruptcy risk and performance increases from their engagement of entrusted loan businesses in the year the loans are issued and in the following year. Further, firms’ risk increases and performance improves significantly when firms are financially healthy, financially constrained, and non-state-owned. Overall, our findings provide policy implications that the risk of shadow banking activities must be cautious.

TGF-β1 induces VEGF expression in human granulosa-lutein cells: a potential mechanism for the pathogenesis of ovarian hyperstimulation syndrome

Lanlan Fang,Yiran Li,Sijia Wang,Yuxi Li,Hsun-Ming Chang,Yuyin Yi,Yang-yang Sun,Avinash Thakur,Peter C. K. Leung,Jung-Chien Cheng,Ying-Pu Sun 생화학분자생물학회 2020 Experimental and molecular medicine Vol.52 No.-

Ovarian hyperstimulation syndrome (OHSS) is one of the most serious and iatrogenic complications that can occur during in vitro fertilization treatment. Although the pathogenesis of OHSS is not fully understood, vascular endothelial growth factor (VEGF) has been recognized as an important mediator of the development of OHSS. Transforming growth factor-beta-1 (TGF-β1) is known to regulate various ovarian functions. However, whether VEGF can be regulated by TGF-β1 in human granulosa cells has not been determined. In addition, the role of TGF-β1 in the pathogenesis of OHSS remains unknown. In the present study, we demonstrate that TGF-β1 stimulates VEGF expression in and secretion from both immortalized human granulosa-lutein (hGL) cells and primary hGL cells. Our results demonstrate that the SMAD2/3, ERK1/2, and p38 MAPK signaling pathways are involved in TGF-β1-induced VEGF expression and secretion. Using a mouse OHSS model, we show that the expression levels of TGF-β1 and VEGF are increased in the ovaries of OHSS mice. Blocking TGF-β1 signaling inhibits the development of OHSS by attenuating VEGF expression. Moreover, clinical results reveal that the protein levels of TGF-β1 and VEGF are increased in the follicular fluid of patients with OHSS, and that the levels of these two proteins in the follicular fluid are positively correlated. The results of this study help to elucidate the mechanisms by which VEGF expression is regulated in hGL cells, which could lead to the development of alternative therapeutic approaches for treating OHSS.

Protein from Hylocereus polyrhizus protects MRC-5 cells against hydrogen peroxide (H2O2)-induced damage

Haomiao Ding,Yuzhe Zhang,Yue Zha,Sijia Zhou,Chaoqing Cao,Huajian Zhu,Feng Xu,Xiuqiang Zhang,Caisheng Wang 한국식품과학회 2022 Food Science and Biotechnology Vol.31 No.13

The cytoprotective and potential molecular mechanisms of Hylocereus polyrhizus protein (RFPP) were investigated on the hydrogen peroxide (H2O2)-triggered damage in normal human embryonic lung (MRC-5) cells. An MTT assay was conducted to assess the MRC-5 cell viability after exposure to H2O2 or RFPP. Cell cycle distribution and apoptosis were explored via flow cytometry. The contents of related proteins were assessed via western blot. MRC-5 cells exhibited markedly decreased cellular viability after treatment with H2O2; however, treatment with RFPP suppressed this decrease. Additionally, RFPP interference dampened H2O2-triggered intracellular apoptosis levels and increased H2O2-triggered intracellular S phase. In these processes, the contents of phosphorylated (p)-AKT along with p-mTOR proteins were downregulated in 120 µM H2O2-treated cells compared with vehicle-treated cells. Nevertheless, in MRC-5 cells inoculated with RFPP, the levels expression of these proteins were reversed. To conclude, RFPP protected MRC-5 cells from H2O2-triggered damage via activation of the PI3K/AKT/mTOR cascade.

Effect of Aged Lubricating Oil on the Regeneration of Diesel Particulate Filters and Ash Physical Characteristics with Non-Thermal Plasma Technology

Zhao Nan,Cai Yi-xi,Shi Yunxi,Wang Weikai,Ni Sijia 한국자동차공학회 2021 International journal of automotive technology Vol.22 No.5

The objective of this study was to evaluate the effects of lubricating oil age on the generation of diesel particulate filters (DPF) by non-thermal plasma (NTP) technology and to characterize the physical properties of ash. The regeneration status was evaluated by the concentration of regeneration products and regeneration temperature. The compositional and morphological characteristics of ash were analyzed by energy-dispersive X-ray spectroscopy (EDX), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Compared with DPF@L-fresh, the internal regeneration temperature was reduced in DPF@L-age. The unit removal time of carbon was 39.23 min/g for DPF@L-fresh but was reduced to 34.87 min/g for DPF@L-age, which indicated that the efficiency of NTP regeneration increased. Shorter regeneration time and lower airflow resistance caused by NTP technology aided the formation of a unique chain-type ash with the structure of a hollow column. Several fine particles that had formed by the condensation of volatiles were distributed on the ash surface of the aged lubricant, and the ash clusters were loosely combined. TEM images revealed that ash was mostly composed of a crystal structure, that the ash dimension of aged lubricant decreased, and that the adhesion between ash particulates was weak.