Enhancement of Aggregation-Induced Emission in Dye-Encapsulating Polymeric Micelles for Bioimaging

Wu, Wen-Chung,Chen, Ching-Yi,Tian, Yanqing,Jang, Sei-Hum,Hong, Yuning,Liu, Yang,Hu, Rongrong,Tang, Ben Zhong,Lee, Yi-Ting,Chen, Chin-Ti,Chen, Wen-Chang,Jen, Alex K.-Y. WILEY-VCH Verlag 2010 Advanced Functional Materials Vol.20 No.9

<P>Three amphiphilic block copolymers are employed to form polymeric micelles and function as nanocarriers to disperse hydrophobic aggregation-induced emission (AIE) dyes, 1,1,2,3,4,5-hexaphenylsilole (HPS) and/or bis(4-(N-(1-naphthyl) phenylamino)-phenyl)fumaronitrile (NPAFN), into aqueous solution for biological studies. Compared to their virtually non-emissive properties in organic solutions, the fluorescence intensity of these AIE dyes has increased significantly due to the spatial confinement that restricts intramolecular rotation of these dyes and their better compatibility in the hydrophobic core of polymeric micelles. The effect of the chemical structure of micelle cores on the photophysical properties of AIE dyes are investigated, and the fluorescence resonance energy transfer (FRET) from the green-emitting donor (HPS) to the red-emitting acceptor (NPAFN) is explored by co-encapsulating this FRET pair in the same micelle core. The highest fluorescence quantum yield (∼62%) could be achieved by encapsulating HPS aggregates in the micelles. Efficient energy transfer (>99%) and high amplification of emission (as high as 8 times) from the NPAFN acceptor could also be achieved by spatially confining the HPS/NPAFN FRET pair in the hydrophobic core of polymeric micelles. These micelles could be successfully internalized into the RAW 264.7 cells to demonstrate high-quality fluorescent images and cell viability due to improved quantum yield and reduced cytotoxicity.</P> <B>Graphic Abstract</B> <P>Highly efficient fluorescence probes are achieved through the encapsulation of aggregation-induced emission molecules, 1,1,2,3,4,5-hexaphenylsilole (HPS) and/or bis(4-(N-(1-naphthyl) phenylamino)-phenyl)fumaronitrile (NPAFN) in the core of polymeric micelles. Bright fluorescence cell images are shown with tunable colors of green directly from HPS aggregates and red through efficient fluorescence resonance energy transfer (FRET) from HPS aggregates to NPAFN aggregates. <img src='wiley_img_2010/1616301X-2010-20-9-ADFM200902043-content.gif' alt='wiley_img_2010/1616301X-2010-20-9-ADFM200902043-content'> </P>

Involvement of Lysosome Membrane Permeabilization and Reactive Oxygen Species Production in the Necrosis Induced by Chlamydia muridarum Infection in L929 Cells

( Lixiang Chen ),( Cong Wang ),( Shun Li ),( Xin Yu ),( Xue Liu ),( Rongrong Ren ),( Wenwen Liu ),( Xiaojing Zhou ),( Xiaonan Zhang ),( Xiaohui Zhou ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.4

Chlamydiae, obligate intracellular bacteria, are associated with a variety of human diseases. The chlamydial life cycle undergoes a biphasic development: replicative reticulate bodies (RBs) phase and infectious elementary bodies (EBs) phase. At the end of the chlamydial intracellular life cycle, EBs have to be released to the surrounded cells. Therefore, the interactions between Chlamydiae and cell death pathways could greatly influence the outcomes of Chlamydia infection. However, the underlying molecular mechanisms remain elusive. Here, we investigated host cell death after Chlamydia infection in vitro, in L929 cells, and showed that Chlamydia infection induces cell necrosis, as detected by the propidium iodide (PI)-Annexin V double-staining flow-cytometric assay and Lactate dehydrogenase (LDH) release assay. The production of reactive oxygen species (ROS), an important factor in induction of necrosis, was increased after Chlamydia infection, and inhibition of ROS with specific pharmacological inhibitors, diphenylene iodonium (DPI) or butylated hydroxyanisole (BHA), led to significant suppression of necrosis. Interestingly, live-cell imaging revealed that Chlamydia infection induced lysosome membrane permeabilization (LMP). When an inhibitor upstream of LMP, CA-074-Me, was added to cells, the production of ROS was reduced with concomitant inhibition of necrosis. Taken together, our results indicate that Chlamydia infection elicits the production of ROS, which is dependent on LMP at least partially, followed by induction of host-cell necrosis. To our best knowledge, this is the first live-cell-imaging observation of LMP post Chlamydia infection and report on the link of LMP to ROS to necrosis during Chlamydia infection.

Detection for Operation Chain: Histogram Equalization and Dither-like Operation

( Zhipeng Chen ),( Yao Zhao ),( Rongrong Ni ) 한국인터넷정보학회 2015 KSII Transactions on Internet and Information Syst Vol.9 No.9

Many sorts of image processing software facilitate image editing and also generate a great number of doctored images. Forensic technology emerges to detect the unintentional or malicious image operations. Most of forensic methods focus on the detection of single operations. However, a series of operations may be used to sequentially manipulate an image, which makes the operation detection problem complex. Forensic investigators always want to know as much exhaustive information about a suspicious image`s entire processing history as possible. The detection of the operation chain, consisting of a series of operations, is a significant and challenging problem in the research field of forensics. In this paper, based on the histogram distribution uniformity of a manipulated image, we propose an operation chain detection scheme to identify histogram equalization (HE) followed by the dither-like operation (DLO). Two histogram features and a local spatial feature are utilized to further determine which DLO may have been applied. Both theoretical analysis and experimental results verify the effectiveness of our proposed scheme for both global and local scenarios.

Average Modeling and Control of Module Multilevel Converter

Zhang, GuoJu,Chen, Yao,Qi, Lisa,Yu, Rongrong,Pan, Jiuping The Korean Institute of Electrical Engineers 2014 The Journal of International Council on Electrical Vol.4 No.2

Modular Multilevel Converter (MMC) has presents great potential in high power quality, low operation loss, scalability and high reliability, which make MMC a good choice for DC transmission, MV drive and other HV or MV applications. This paper describes the average modeling and control of MMC. Firstly, the operation principles of MMC are analyzed, based on which the average MMC model is deduced. Secondly, the control methods for DC voltage control, circulating current suppression, and capacitor voltage balancing, etc are developed. Finally, the effectiveness of the proposed average model is verified by comparing the simulation results of an MVDC distribution system using detailed MMC models. The correctness of the designed control methods also demonstrated through simulation. It is also proven to be feasible of using MMC in MVDC applications.

Genomic Characteristics and the Potential Clinical Implications in Oligometastatic Non–Small Cell Lung Cancer

Rongxin Liao,Kehong Chen,Jinjin Li,Hengqiu He,Guangming Yi,Mingfeng Huang,Rongrong Chen,Lu Shen,Xiaoyue Zhang,Zaicheng Xu,Zhenzhou Yang,Yuan Peng 대한암학회 2023 Cancer Research and Treatment Vol.55 No.3

Purpose Oligometastatic non–small cell lung cancer (NSCLC) patients have been increasingly regarded as a distinct group that could benefit from local treatment to achieve a better clinical outcome. However, current definitions of oligometastasis are solely numerical, which are imprecise because of ignoring the biological heterogeneity caused by genomic characteristics. Our study aimed to profile the molecular alterations of oligometastatic NSCLC and elucidate its potential difference from polymetastasis. Materials and Methods We performed next-generation sequencing to analyze tumors and paired peripheral blood from 77 oligometastatic and 21 polymetastatic NSCLC patients to reveal their genomic characteristics and assess the genetic heterogeneity. Results We found ERBB2, ALK, MLL4, PIK3CB, and TOP2A were mutated at a significantly lower frequency in oligometastasis compared with polymetastasis. EGFR and KEAP1 alterations were mutually exclusive in oligometastatic group. More importantly, oligometastasis has a unique significant enrichment of apoptosis signaling pathway. In contrast to polymetastasis, a highly enriched COSMIC signature 4 and a special mutational process, COSMIC signature 14, were observed in the oligometastatic cohort. According to OncoKB database, 74.03% of oligometastatic NSCLC patients harbored at least one actionable alteration. The median tumor mutation burden of oligometastasis was 5.00 mutations/Mb, which was significantly associated with smoking, DNA damage repair genes, TP53 mutation, SMARCA4 mutation, LRP1B mutation, ABL1 mutation. Conclusion Our results shall help redefine oligometastasis beyond simple lesion enumeration that will ultimately improve the selection of patients with real oligometastatic state and optimize personalized cancer therapy for oligometastatic NSCLC.

High yield engineered nanovesicles from ADSC with enriched miR-21-5p promote angiogenesis in adipose tissue regeneration

Sun Di,Mou Shan,Chen Lifeng,Yang Jie,Wang Rongrong,Zhong Aimei,Wang Wei,Tong Jing,Wang Zhenxing,Sun Jiaming 한국생체재료학회 2023 생체재료학회지 Vol.27 No.00

Background: Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) have been found to have a great potential for soft tissue repair due to various biological functions, including pro-angiogenesis and low immunogenicity. However, the low yield and heterogeneity of MSC-EVs limited their clinical transformation. This study was designed to develop a novel adipose-derived stem cell engineered nanovesicles (ADSC-NVs) with high production and explore its pro-angiogenetic effect and application in adipose tissue regeneration. Methods: Adipose-derived stem cell-derived extracellular vesicles (ADSC-EVs) were isolated from an EVs-free culture medium for human ADSCs (hADSCs). ADSC-NVs were prepared by sequentially extruding ADSCs followed by iodixanol density gradient ultracentrifugation and were compared with ADSC-EVs in morphology, size distribution, protein contents and yield. The pro-angiogenetic effect of ADSC-NVs in different doses (0, 5, 20 and 80 μg/mL) in vitro was determined using transwell assay, tube formation assay, western blot and qRT-PCR. In vivo, BALB/c nude mice were administered injection of a mixture of fat granules and different dose of ADSC-NVs and grafts were harvested at 12 weeks post-transplantation for further analysis. By analyzing the weight and volume of grafts and histological evaluation, we investigated the effect of ADSC-NVs in vessel formation and adipose tissue regeneration. Results: Our results showed yield of purified ADSC-NVs was approximately 20 times more than that of ADSC-EVs secreted by the same number of ADSCs. In vitro, both ADSC-NVs and ADSC-EVs exhibited a dose-dependent proangiogenetic effect, despite their distinct miRNA profiles. These effects of ADSC-NVs may be mediated by enriched miR-21-5p via PTEN inhibition and PI3K/p-Akt signaling activation. Furthermore, after a mixed injection of ADSC-NVs, vessel formation and adipose regeneration were observed in vivo in fat implants. Conclusions: Our study developed a potent alternative of ADSC-EVs. ADSC-NVs have a high pro-angiogenesis potential and can be used as cell-free therapeutic biomaterials in soft tissue regeneration.

Influence of Simulated Precipitation on Dryland Soil Respiration in the Loess Plateau, China

Jun Wang,Quanquan Liu,Rongrong Chen,Upendra M. Sainju 한국토양비료학회 2014 한국토양비료학회 학술발표회 초록집 Vol.2014 No.6

A core–shell polymeric–inorganic hybrid nanocomposite system for MRI-visible gene delivery application in cancer immunotherapy

Jiali Cai,Guochuang Chen,Rongrong Jin,Changhui Deng,Shihui Huang,Xuexia Yuan,Gengjia Chen,Jing Zhao,Zhiyong Wang,Hua Ai 한국공업화학회 2019 Journal of Industrial and Engineering Chemistry Vol.76 No.-

Due to the tumor immune escape mechanism, strategies like the utilization of bispecific antibodies wereused to assist immunotherapy. Herein a visible polymer–inorganic hybrid gene vehicle had beenprepared to deliver anti-EpCAM/anti-CD3 encoding minicircle DNA into normal cells thus thesetransfected cells effectively secreted bispecific antibody with bioactivity. In the experiments, thiscompound system exhibited excellent biocompatibility, high transfection efficiency and ultrasensitiveimaging capacity. Furthermore, the transfected cells could be detected under the MR imaging system andthe gene product showed an outstanding immune effect. In conclusion, this nanocomposite showedsynergistic advantages in gene delivery and non-invasive MR imaging.

Genomic and Transcriptomic Characterization Revealed the High Sensitivity of Targeted Therapy and Immunotherapy in a Subset of Endometrial Stromal Sarcoma

Nan Kang,Yinli Zhang,Shichao Guo,Ran Chen,Fangzhou Kong,Shuchun Wang,Mingming Yuan,Rongrong Chen,Danhua Shen,Jianliu Wang 대한암학회 2023 Cancer Research and Treatment Vol.55 No.3

Purpose The unique chromosomal rearrangements of endometrial stromal sarcoma (ESS) make it possible to distinguish high-grade ESS (HGESS) and low-grade ESS (LGESS) from the molecular perspective. Analysis of ESS at the genomic and transcriptomic levels can help us achieve accurate diagnosis of ESS and provide potential therapy options for ESS patients.Materials and Methods A total of 36 ESS patients who conducted DNA- and/or RNA-based next-generation sequencing were retrospectively enrolled in this study. The molecular characteristics of ESS at genomic and transcriptomic levels, including mutational spectrum, fusion profiles, gene expression and pathway enrichment analysis and features about immune microenvironment were comprehensively explored.Results <i>TP53</i> and <i>DNMT3A</i> mutations were the most frequent mutations. The classical fusions frequently found in <i>HGESS</i> (<i>ZC3H7B-BCOR</i> and <i>NUTM2B-YWHAE</i>) and LGESS (<i>JAZF1-SUZ12</i>) were detected in our cohort. <i>CCND1</i> was significantly up-regulated in HGESS, while the expression of <i>GPER1</i> and <i>PGR</i> encoding estrogen receptor (ER) and progesterone receptor (PR) did not differ significantly between HGESS and LGESS. Actionable mutations enriched in homologous recombination repair, cell cycle, and phosphoinositide 3-kinase/AKT/mammalian target of rapamycin pathways were detected in 60% of HGESS patients. Genes with up-regulated expression in HGESS were significantly enriched in five immune-related pathways. Most HGESS patients (85.7%) had positive predictors of immunotherapy efficacy. Moreover, immune microenvironment analysis showed that HGESS had relatively high immune infiltration. The degree of immune infiltration in HGESS patients with <i>ZC3H7B-BCOR</i> fusion was relatively higher than that of those with <i>NUTM2B-YWHAE</i> fusion.Conclusion This study investigated the molecular characteristics of ESS patients at the genomic and transcriptomic levels and revealed the potentially high sensitivity of targeted therapy and immunotherapy in a subset of HGESS with specific molecular features, providing a basis for guiding decision-making of treatment and the design of future clinical trials on precision therapy.

CDH17 nanobodies facilitate rapid imaging of gastric cancer and efficient delivery of immunotoxin

Jingbo Ma,Xiaolong Xu,Chunjin Fu,Peng Xia,Ming Tian,Liuhai Zheng,Kun Chen,Xiaolian Liu,Yilei Li,Le Yu,Qinchang Zhu,Yangyang Yu,Rongrong Fan,Haibo Jiang,Zhifen Li,Chuanbin Yang,Chengchao Xu,Ying Long,J 한국생체재료학회 2022 생체재료학회지 Vol.26 No.4

Background: It is highly desirable to develop new therapeutic strategies for gastric cancer given the low survival rate despite improvement in the past decades. Cadherin 17 (CDH17) is a membrane protein highly expressed in cancers of digestive system. Nanobody represents a novel antibody format for cancer targeted imaging and drug delivery. Nanobody targeting CHD17 as an imaging probe and a delivery vehicle of toxin remains to be explored for its theragnostic potential in gastric cancer. Methods: Naïve nanobody phage library was screened against CDH17 Domain 1-3 and identified nanobodies were extensively characterized with various assays. Nanobodies labeled with imaging probe were tested in vitro and in vivo for gastric cancer detection. A CDH17 Nanobody fused with toxin PE38 was evaluated for gastric cancer inhibition in vitro and in vivo. Results: Two nanobodies (A1 and E8) against human CDH17 with high affinity and high specificity were successfully obtained. These nanobodies could specifically bind to CDH17 protein and CDH17-positive gastric cancer cells. E8 nanobody as a lead was extensively determined for tumor imaging and drug delivery. It could efficiently co-localize with CDH17-positive gastric cancer cells in zebrafish embryos and rapidly visualize the tumor mass in mice within 3 h when conjugated with imaging dyes. E8 nanobody fused with toxin PE38 showed excellent anti-tumor effect and remarkably improved the mice survival in cell-derived (CDX) and patient-derived xenograft (PDX) models. The immunotoxin also enhanced the anti-tumor effect of clinical drug 5-Fluorouracil. Conclusions: The study presents a novel imaging and drug delivery strategy by targeting CDH17. CDH17 nanobodybased immunotoxin is potentially a promising therapeutic modality for clinical translation against gastric cancer.