Cellular uptake mechanism and comparative in vitro cytotoxicity studies of monomeric LMWP-siRNA conjugate

Junxiao Ye,Xing Pei,Hui Cui,Zhili Yu,이혁진,Jian-Xin Wang,Xu Wang,Lu Sun,Huining He,Victor C. Yang 한국공업화학회 2018 Journal of Industrial and Engineering Chemistry Vol.63 No.-

The covalent attachment of CPPs to siRNA molecules offers great potential for CPP-mediated siRNA delivery. We recently reported a concise and high-yield synthesis strategy of the cell-permeable, cytosol-dissociable LMWP-siRNA covalent conjugate. Herein, cell uptake mechanism and cellular toxicity studies of this conjugate were performed to evaluate the potential of LMWP-siRNA conjugate for clinical translation. Cellular uptake mechanism study indicated that the conjugate could be taken up by cells via multiple pathways, including direct penetration of the plasma membrane and clathrin- and caveolae-independent endocytosis. In vitro cytotoxicity study revealed that the conjugation promoted internalization in a low-toxic fashion.

Cellular uptake mechanism and comparative <i>in vitro</i> cytotoxicity studies of monomeric LMWP-siRNA conjugate

Ye, Junxiao,Pei, Xing,Cui, Hui,Yu, Zhili,Lee, Hyukjin,Wang, Jianxin,Wang, Xu,Sun, Lu,He, Huining,Yang, Victor C. THE KOREAN SOCIETY OF INDUSTRIAL AND ENGINEERING 2018 JOURNAL OF INDUSTRIAL AND ENGINEERING CHEMISTRY -S Vol.63 No.-

<P><B>Abstract</B></P> <P>The covalent attachment of CPPs to siRNA molecules offers great potential for CPP-mediated siRNA delivery. We recently reported a concise and high-yield synthesis strategy of the cell-permeable, cytosol-dissociable LMWP-siRNA covalent conjugate. Herein, cell uptake mechanism and cellular toxicity studies of this conjugate were performed to evaluate the potential of LMWP-siRNA conjugate for clinical translation. Cellular uptake mechanism study indicated that the conjugate could be taken up by cells via multiple pathways, including direct penetration of the plasma membrane and clathrin- and caveolae-independent endocytosis. <I>In vitro</I> cytotoxicity study revealed that the conjugation promoted internalization in a low-toxic fashion.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Conjugated Polymers Based on Modified Benzo[1,2-b:4,5-b']Dithiophene and Perylene Diimide Derivatives: Good Optical Properties and Tunable Electrochemical Performance

Kun Fang,Xu Ye,Yawen Huang,Guanjun Chang,Junxiao Yang,Yueqin Shen 한국고분자학회 2015 Macromolecular Research Vol.23 No.6

Three original conjugated polymers were devised and synthesized via the alternating conjugated polymerization of the electron-donating monomer of the modified benzo[1,2-b:4,5-b']dithiophene (BDT) and electronaccepting monomers of perylene diimide (PDI), dithienyl-perylene diimide (DTPDI) and dithienocoronene diimide (DTCDI), respectively. The synthesized conjugated polymers presented excellent thermal stability (Td >400 oC) as well as broad absorption in visible region and narrow gap of energy level (1.80 to 2.01 eV). Moreover, P(BDT-DTCDI) exhibited strong fluorescence properties ( =736 nm with excitation at 680 nm). According to the theoretical calculations, P(BDT-PDI) and P(BDT-DTPDI) displayed significant polymer backbone torsion (41.44o and 39.64o), and P(BDT-DTCDI) had highly coplanar backbone with negligible angle between BDT and DTCDI. The results indicated that P(BDT-DTCDI) had potential applications in the fields of organic thin-film transistors (OTFTs) and fluorescence materials, and P(BDT-PDI) and P(BDT-DTPDI) could be used as a stable n-type semiconductor for polymer solar cells.

De novo transcriptomic analysis of gonad of Strongylocentrotus nudus and gene discovery for biosynthesis of polyunsaturated fatty acids

Zhenlin Wei,Xiaolin Liu,Zunchun Zhou,Junxiao Xu 한국유전학회 2019 Genes & Genomics Vol.41 No.5

Background Strongylocentrotus nudus is an important cultured sea urchin species in north China, because its gonad is rich in unsaturated fatty acids, particularly long polyunsaturated fatty acids (LC-PUFAs). These PUFAs play pleiotropic and crucial roles in a wide range of biological process. Objective However, the genes contributing to biosynthesis PUFAs have not been elucidated yet, and the molecular mechanism relative to the difference in PUFA composition between male and female gonad as been revealed but the corresponding has not been understood. Methods In this paper, solexa sequencing based transcriptomic approach was used to identify and characterize the key genes relative to PUFA synthesis and further conducted different expressed genes between male and female gonad. Results A total of 130,124 transcripts and 189330 unigenes were de novo assembled from 64.32 Gb data. Next, these unigenes were subjected to functional annotation by mapping to six public databases, and this process revealed a lot of genes involving in lipid metabolism. In addition, three types of fatty acids front-end desaturase and three species of very long fatty acids elongase were identified and the pathway for PUFA biosynthesis was hypothesized. Last, comparative analysis revealed the higher expression level of Δ5 desaturase, Δ6 desaturase, ELOVL-4, -6 and -7 in male gonad compared with female. Conclusion This results could plausible explain the differ in composition of PUFAs between male and female gonad of sea urchin.

All-Benzocyclobutene Functionalized Polycarbosilane and Derived Copolymers with Low Dielectric Constant and High Thermal Stability

Nan Zhong,Xian Li,Huan Hu,Yawen Huang,Xu Ye,Junxiao Yang 한국고분자학회 2019 Macromolecular Research Vol.27 No.12

All-benzocyclobutene functionalized polycarbosilanes and the derived copolymers were successfully synthesized by ring-opening polymerization (ROP) of 1,1-dibenzocyclobutene-1-silacyclobutane (DBCBSCB) and 1,1-diphenylsiletane (DPSCB) using Karstedt’s catalyst. Adjusting the copolymerization ratio of DBCBSCB and DPSCB could afford to control the ultimate properties of the cured resins. The linear polycarbosilanes containing dibenzocyclobutene were directly cross-linked upon high temperature. With increasing the copolymerization ratio of DBCBSCB and DPSCB, the 5% weight loss temperature (T5%) of cured resins is increased from 450 °C to 506 °C under a nitrogen atmosphere, while the dielectric constant (k) is decreased from 2.60 to 2.42 at 10 MHz. The excellent thermostability and low-dielectric performance make such thermosets have potential application in the microelectronic industry.

pH and Redox Dual Stimuli-Responsive Injectable Hydrogels Based on Carboxymethyl Cellulose Derivatives

Yueqin Shen,Xian Li,Yawen Huang,Guanjun Chang,Ke Cao,Junxiao Yang,Renyi Zhang,Xueying Sheng,Xu Ye 한국고분자학회 2016 Macromolecular Research Vol.24 No.7

pH and redox dual stimuli-responsive injectable hydrogels were prepared by cross-linking oxidized carboxymethyl cellulose (oxi-CMC) with 3,3'-dithiobis (propionohydrazide) (DTP) via Schiff base reaction under physiological condition. The hydrogels showed good performance such as tunable gelling time, appropriate rheology properties, high swelling ratio and low degradation rate. In vitro release studies confirmed that bovine serum albumin (BSA) as a model drug exhibited a sustainable release at pH 7.4 and an accelerated release under a lower pH 5.0 and/or reducing environments. The results signified that oxi-CMC/DTP hydrogels could be an attractive candidate for drug delivery system, tissue engineering or cell scaffold materials.

Gene expression and promoter methylation of porcine uncoupling protein 3 gene

Ruiyi Lin,Weimin Lin,Qiaohui Chen,Jianchao Huo,Yuping Hu,Junxiao Ye,Jingya Xu,Tianfang Xiao 아세아·태평양축산학회 2019 Animal Bioscience Vol.32 No.2

Objective: Uncoupling protein 3 gene (UCP3) is a candidate gene associated with the meat quality of pigs. The aim of this study was to explore the regulation mechanism of UCP3 expression and provide a theoretical basis for the research of the function of porcine UCP3 gene in meat quality. Methods: Bisulfite sequencing polymerase chain reaction (PCR) and quantitative real-time PCR (Q-PCR) were used to analyze the methylation of UCP3 5′-flanking region and UCP3 mRNA expression in the adipose tissue or skeletal muscle of three pig breeds at different ages (1, 90, 210-day-old Putian Black pig; 90-day-old Duroc; and 90-day-old Dupu). Results: Results showed that two cytosine-guanine dinucleotide (CpG) islands are present in the promoter region of porcine UCP3 gene. The second CpG island located in the core promoter region contained 9 CpG sites. The methylation level of CpG island 2 was lower in the adipose tissue and skeletal muscle of 90-day-old Putian Black pigs compared with 1-day-old and 210-day-old Putian Black pigs, and the difference also existed in the skeletal muscle among the three 90-day-old pig breeds. Furthermore, the obvious changing difference of UCP3 mRNA expression was observed in the skeletal muscle of different groups. However, the difference of methylation status and expression level of UCP3 gene was not significant in the adipose tissue. Conclusion: Our data indicate that UCP3 mRNA expression level was associated with the methylation status of UCP3 promoter in the skeletal muscle of pigs.