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      • Modulation of Drug Resistance in Ovarian Cancer Cells by Inhibition of Protein Kinase C-alpha (PKC-α) with Small Interference RNA (siRNA) Agents

        Zhao, Li-Jun,Xu, Heng,Qu, Jun-Wei,Zhao, Wan-Zhou,Zhao, Yi-Bing,Wang, Jin-Hua Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.8

        Objective: To determine whether silence of $PKC-{\alpha}$ expression by small interference RNA (siRNA) might regulate MDR1 expression and reverse chemoresistance of ovarian cancer. Methods: We measured gene and protein expression of MDR1 and $PKC-{\alpha}$ in ovarian cancer cells and assessed their correlation with cell drug resistance. We also examined whether blocking $PKC-{\alpha}$ by RNA interference (RNAi) affected MDR1 expression and reversed drug resistance in drug sensitivity tests. Results: The drug resistance cell lines, OV1228/DDP and OV1228/Taxol, had higher gene and protein expression of MDR1 and $PKC-{\alpha}$ than their counterpart sensitive cell line, OV1228. SiRNA depressed $PKC-{\alpha}$ gene protein expression, as well as MDR1 and protein expression and improved the drug sensitivity in OV1228/DDP and OV1228/Taxol cells. Conclusion: These results indicated that decreasing $PKC-{\alpha}$ expression with siRNA might be an effective method to improve drug sensitivity in drug resistant cells with elevated levels of $PKC-{\alpha}$ and MDR1. A new siRNA-based therapeutic strategy targeting $PKC-{\alpha}$ gene could be designed to overcome the chemoresistance of ovarian cancer.

      • Effects of polymer surface energy on morphology and properties of silver nanowire fabricated via nanoimprint and E-beam evaporation

        Zhao, Zhi-Jun,Hwang, Soon Hyoung,Jeon, Sohee,Jung, Joo-Yun,Lee, Jihye,Choi, Dae-Geun,Choi, Jun-Hyuk,Park, Sang-Hu,Jeong, Jun-Ho Elsevier 2017 APPLIED SURFACE SCIENCE - Vol.420 No.-

        <P><B>Abstract</B></P> <P>In this paper, we demonstrate that use of different nanoimprint resins as a polymer pattern has a significant effect on the morphology of silver (Ag) nanowires deposited via an E-beam evaporator. RM-311 and Ormo-stamp resins are chosen as a polymer pattern to form a line with dimensions of width (100nm)×space (100nm)×height (120nm) by using nanoimprint lithography (NIL). Their contact angles are then measured to evaluate their surface energies. In order to compare the properties of the Ag nanowires deposited on the various polymer patterns with different surface energies, hydrophobic surface treatment of the polymer pattern surface is implemented using self-assembled monolayers. In addition, gold and aluminum nanowires are fabricated for comparison with the Ag nanowires, with the differences in the nanowire morphologies being determined by the different atomic properties. The monocrystalline and polycrystalline structures of the various Ag nanowire formations are observed using transmission electron microscopy. In addition, the melting temperatures and optical properties of four kinds of Ag nanowire morphologies deposited on various polymer patterns are evaluated using a hot plate and an ultraviolet-visible (UV–vis) spectrometer, respectively. The results indicate that the morphology of the Ag nanowire determines the melting temperature and the transmission. We believe that these findings will greatly aid the development of NIL, along with physical evaporation and chemical deposition techniques, and will be widely employed in optics, biology, and surface wettability applications.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Ag nanowires with various morphologies deposited on polymer patterns with different surface energies were fabricated via nanoimprint and e-beam evaporation. </LI> <LI> The formation mechanism behind the different morphologies of the Ag nanowires deposited on the polymer patterns with different surface energies was explained and confirmed. </LI> <LI> The morphologies of Ag nanowires have an important effect on their transmission and melting temperature. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

      • Macrophages Promote Coal Tar Pitch Extract-induced Tumorigenesis of BEAS-2B Cells and Tumor Metastasis in Nude Mice Mediated by AP-1

        Zhang, Peng,Jin, Yue-Fei,Zhang, Qiao,Wu, Yi-Ming,Wu, Wei-Dong,Yao, Wu,Wu, Yong-Jun,Li, Zhi-Tao,Zhao, Yong,Liu, Yu,Feng, Fei-Fei Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.12

        Background: We sought to evaluate the role of tumor associated macrophages (TAMs) on the promotion of coal tar pitch extract (CTPE)-induced tumorigenesis of human bronchial epithelial cells (BEAS-2B) and tumor metastasis in nude mice, and related mechanisms. Materials and Methods: BEAS-2B cells were first treated with 2.4 mg/mL CTPE for 72 hours. After removal of CTPE, the cells were continuously cultured and passaged using trypsin-EDTA. THP-1 cells were used as macrophage-like cells. BEAS-2B cells under different conditions (n=6/group) were injected into the back necks of nude mice, and alterations of tumor xenograft growth, indicative of tumorigenicity, and tumor metastasis were determined. Pathological changes (tumor nests and microvascular lesions) of HE-stained tumor tissues were also evaluated. The expression of AP-1(c-Jun) in xenografts and metastatic tumors was determined using immunohistochemistry. Results: Tumor size and weight in nude mice transplanted with the mixture of CTPE-induced passage 30 BEAS-2B and THP-1 cells (2:1) were increased compared to those from the CTPE-treated BEAS-2B cells at passage 30 alone at different observation time points. Tumor metastasis to lymph nodes and liver was only detected after transplantation of a mixture the two kinds of cells. The numbers of tumor nests and microvascular lesions, and the expression levels of AP-1 (c-Jun) in tumors from the mixture of two kinds of cells were increased apparently in contrast to those in tumor from the CTPE-treated BEAS-2B cells of passage 30 alone. In addition, there was positive correlation between AP-1 (c-Jun) expression level and the number of microvascular lesions, or between AP-1 (c-Jun) expression level and tumor metastasis in these two groups. Conclusions: TAMs not only facilitate tumorigenesis transformation of CTPE-induced BEAS-2B cells, but also promote tumor growth, angiogenesis and metastasis in nude mice in vivo, which may be mediated by AP-1.

      • Optimization of Reference Genes for Normalization of the Quantitative Polymerase Chain Reaction in Tissue Samples of Gastric Cancer

        Zhao, Lian-Mei,Zheng, Zhao-Xu,Zhao, Xiwa,Shi, Juan,Bi, Jian-Jun,Pei, Wei,Feng, Qiang Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.14

        For an exact comparison of mRNA transcription in different samples or tissues with real time quantitative reverse transcription-polymerase chain reaction (qRT-PCR), it is crucial to select a suitable internal reference gene. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and beta-actin (ACTB) have been frequently considered as house-keeping genes to normalize for changes in specific gene expression. However, it has been reported that these genes are unsuitable references in some cases, because their transcription is significantly variable under particular experimental conditions and among tissues. The present study was aimed to investigate which reference genes are most suitable for the study of gastric cancer tissues using qRT-PCR. 50 pairs of gastric cancer and corresponding peritumoral tissues were obtained from patients with gastric cancer. Absolute qRT-PCR was employed to detect the expression of GAPDH, ACTB, RPII and 18sRNA in the gastric cancer samples. Comparing gastric cancer with corresponding peritumoral tissues, GAPDH, ACTB and RPII were obviously upregulated 6.49, 5.0 and 3.68 fold, respectively. Yet 18sRNA had no obvious expression change in gastric cancer tissues and the corresponding peritumoral tissues. The expression of GAPDH, ${\beta}$-actin, RPII and 18sRNA showed no obvious changes in normal gastric epithelial cells compared with gastric cancer cell lines. The carcinoembryonic antigen (CEA), a widely used clinical tumor marker, was used as a validation gene. Only when 18sRNA was used as the normalizing gene was CEA obviously elevated in gastric cancer tissues compared with peritumoral tissues. Our data show that 18sRNA is stably expressed in gastric cancer samples and corresponding peritumoral tissues. These observations confirm that there is no universal reference gene and underline the importance of specific optimization of potential reference genes for any experimental condition.

      • SCIESCOPUS

        Unconfined compressive strength of PET waste-mixed residual soils

        Zhao, Jian-Jun,Lee, Min-Lee,Lim, Siong-Kang,Tanaka, Yasuo Techno-Press 2015 Geomechanics & engineering Vol.8 No.1

        Plastic wastes, particularly polyethylene terephthalate (PET) generated from used bottled water constitute a worldwide environmental issue. Reusing the PET waste for geotechnical applications not only reduces environmental burdens of handling the waste, but also improves inherent engineering properties of soil. This paper investigated factors affecting shear strength improvement of PET-mixed residual soil. Four variables were considered: (i) plastic content; (ii) plastic slenderness ratio; (iii) plastic size; and (iv) soil particle size. A series of unconfined compression tests were performed to determine the optimum configurations for promoting the shear strength improvement. The results showed that the optimum slenderness ratio and PET content for shear strength improvement were 1:3 and 1.5%, respectively. Large PET pieces (i.e., $1.0cm^2$) were favorable for fine-grained residual soil, while small PET pieces (i.e., $0.5cm^2$) were favorable for coarse-grained residual soil. Higher shear strength improvement was obtained for PET-mixed coarse-grained residual soil (148%) than fine-grained residual soils (117%). The orientation of plastic pieces in soil and frictional resistance developed between soil particles and PET surface are two important factors affecting the shear strength performance of PET-mixed soil.

      • SCIESCOPUSKCI등재

        Production of Transgenic Pigs with an Introduced Missense Mutation of the Bone Morphogenetic Protein Receptor Type IB Gene Related to Prolificacy

        Zhao, Xueyan,Yang, Qiang,Zhao, Kewei,Jiang, Chao,Ren, Dongren,Xu, Pan,He, Xiaofang,Liao, Rongrong,Jiang, Kai,Ma, Junwu,Xiao, Shijun,Ren, Jun,Xing, Yuyun Asian Australasian Association of Animal Productio 2016 Animal Bioscience Vol.29 No.7

        In the last few decades, transgenic animal technology has witnessed an increasingly wide application in animal breeding. Reproductive traits are economically important to the pig industry. It has been shown that the bone morphogenetic protein receptor type IB (BMPR1B) A746G polymorphism is responsible for the fertility in sheep. However, this causal mutation exits exclusively in sheep and goat. In this study, we attempted to create transgenic pigs by introducing this mutation with the aim to improve reproductive traits in pigs. We successfully constructed a vector containing porcine BMPR1B coding sequence (CDS) with the mutant G allele of A746G mutation. In total, we obtained 24 cloned male piglets using handmade cloning (HMC) technique, and 12 individuals survived till maturation. A set of polymerase chain reactions indicated that 11 of 12 matured boars were transgene-positive individuals, and that the transgenic vector was most likely disrupted during cloning. Of 11 positive pigs, one (No. 11) lost a part of the terminator region but had the intact promoter and the CDS regions. cDNA sequencing showed that the introduced allele (746G) was expressed in multiple tissues of transgene-positive offspring of No.11. Western blot analysis revealed that BMPR1B protein expression in multiple tissues of transgene-positive $F_1$ piglets was 0.5 to 2-fold higher than that in the transgene-negative siblings. The No. 11 boar showed normal litter size performance as normal pigs from the same breed. Transgene-positive $F_1$ boars produced by No. 11 had higher semen volume, sperm concentration and total sperm per ejaculate than the negative siblings, although the differences did not reached statistical significance. Transgene-positive $F_1$ sows had similar litter size performance to the negative siblings, and more data are needed to adequately assess the litter size performance. In conclusion, we obtained 24 cloned transgenic pigs with the modified porcine BMPR1B CDS using HMC. cDNA sequencing and western blot indicated that the exogenous BMPR1B CDS was successfully expressed in host pigs. The transgenic pigs showed normal litter size performance. However, no significant differences in litter size were found between transgene-positive and negative sows. Our study provides new insight into producing cloned transgenic livestock related to reproductive traits.

      • KCI등재

        Multi-field coupling finite element analysis for determining the influence of temperature field on die service life during precision-forming process of steel synchronizer ring

        Jun Zhao,Shan-Ming Luo,Feng-Qiang Li,Chen-Bing Xu 대한기계학회 2017 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.31 No.7

        Failure analysis shows that increased die temperature caused by severe plastic deformation of material and heat conduction between hot billet and cavity significantly affects the distortion of gear cavity in steel synchronizer ring forging process. The forging process of steel synchronizer ring and die temperature distribution under different forging conditions are analyzed through finite element method. Simulation results show that severe plastic deformation occurs in the gear cavity. The improvement of lubrication condition results in decreased die temperature. When the initial billet temperature is high, the die temperature is also high. Increasing forging speed in a certain range facilitates the die temperature decrease. The distribution of die temperature in synthetic forming technology is more reasonable than that of one step forging. The synthetic forming technology is adopted in production to reduce the effects of severe plastic deformation caused by die temperature. The ejection mechanism and control system of the double disc friction press are improved to reduce the contact time between the hot billet and cavity. Experimental results show that synthetic forming technology is reasonable, and that the die service life is prolonged.

      • SCOPUSKCI등재

        Effects of a mild heat treatment on mouse testicular gene expression and sperm quality

        Zhao, Jun,Zhang, Ying,Hao, Linlin,Wang, Jia,Zhang, Jiabao,Liu, Songcai,Ren, Bingzhong The Korean Society for Integrative Biology 2010 Animal cells and systems Vol.14 No.4

        The decrease in sperm quality under heat stress causes a great loss in animal husbandry production. In order to reveal the mechanism underlying the sperm quality decrease caused by heat stress, we first established a mild heat-treated mouse model. Then, the sperm quality was identified. Further, the testicular proteome profile was mapped and compared with the control using 2D electrophoresis and mass spectrometry. Finally, the differential expressed proteins involved in the heat stress response were identified by real-time PCR and Western blotting. The results showed that heat stress caused a significant reduction in mouse sperm quality (P<0.05). Further, 52 protein spots on the 2D gel were found to differ between the heat-shocked tissues and the control. Of these spots, some repair proteins which might provide some explanation for the influence on sperm quality were found. We then focused on Bag-1, Hsp40, Hsp60 and Hsp70, which were found to be differently expressed after heat shock (P<0.05). Further analysis in this heat-shocked model suggests numerous potential mechanisms for heat shock-induced spermatogenic disorders.

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