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A boronate-based modular assembly nanosystem to block the undesirable crosstalk between hepatic stellate cells and Kupffer cells

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Bioactive Materials 2023年第7期25卷 569-579页

作者： Huizhen Jia Ling Ding Ao Yu Weimin Tang Siyuan Tang Chuhan Zhang David Oupický center for Drug Delivery and nanomedicine Department of Pharmaceutical SciencesCollege of PharmacyUniversity of Nebraska Medical CenterOmahaNE68198USA

Crosstalk between Kupffer cells(KCs)and hepatic stellate cells(HSCs)plays an important role in multiple liver disease conditions,including the formation of liver fibrosis in alcohol-associated liver disease(AALD).Therapeutic targeting of the KC-HSC crosstalk is a prime target for therapeutic ***,a novel modular nanosystem was designed and prepared through the self-assembly utilizing boric acid and catechol interactions to prepare polymers modified with a CXCR4-inhibiting *** polymers were used to encapsulate anti-miR-155 and to block the undesirable crosstalk between HSCs and KCs by downregulating miR-155 expression in KCs with the parallel inhibition of CXCR4 signaling in activated *** combined inhibition of miR-155 and CXCR4 at two different liver cell types achieved improved antifibrosis effects in a mouse model of AALD *** finding highlights the key role that blocking the undesirable crosstalk between HSCs and KCs plays in reversing AALD fibrosis as well as demonstrates a proof-of-concept approach for designing and constructing multifunctional delivery nanosystems using orthogonal functional modules based on the understanding of disease mechanisms.

关键词： Nanoparticles miRNA CXCR4 Liver fibrosis Undesirable crosstalk Modular assembly

Development of fluorinated polyplex nanoemulsions for improved small interfering RNA delivery and cancer therapy

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Nano Research 2018年第7期11卷 3746-3761页

作者： Gang Chen Kaikai Wang Pengkai Wu Yixin Wang Zhanwei Zhou Lifang Yin Minjie Sun David Oupicky State Key Laboratory of Natural Medicines Departmen t of Pharmaceutics China Pharmaceutical University Nanjing 210009 China center for Drug Delivery and nanomedicine Department of Pharmaceutical Sciences University of Nebraska Medical Center Omaha NE 68198 USA

We report the development of a small interfering RNA （siRNA） delivery vector based on cationic perfluorocarbon nanoemulsions. We have prepared perfluorodecalin （PFD） emulsions with a positive surface charge provided by a fluorinated poly（ethylenimine）（F-PEI）. The fluorinated emulsion （F-PEI@PFD） reduced cytotoxicity of F-PEI and demonstrated effective binding with siRNAs to form nanosized emulsion polyplexes. The prepared emulsion polyplexes enhanced cellular uptake and improved endosomal escape of the siRNA. In addition to increased reporter gene silencing in multiple cancer cell lines, when compared with control F-PEI and PEI polyplexes, the siR_NA emulsion polyplexes showed an excellent resistance to serum deactivation and maintained high activity, even in high-serum conditions. The F-PEI@PFD emulsion polyplexes carrying an siRNA to silence the expression of Bcl2 gene induced apoptosis and inhibited tumor growth in a melanoma mouse model in vivo and showed potential for in vivo ultrasound imaging. This study demonstrates the potential of F-PEI@PFD emulsions as a multifunctional theranostic nanoplatform for safe siRNA delivery, with integrated ultrasound imaging functionality.

关键词： polyplexes fluorous interactions small interfering RNA(siRNA) delivery tumor deliver perfluorocarbon emulsion

Application of Phage-displayed Single Chain Antibodies in Western Blot

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Acta Biochimica et Biophysica Sinica 2005年第3期37卷 205-209页

作者： Yan-Li DING Mei-Yun LIU Wei HAN Sheng-Li YANG Hui LIU Yi GONG School of pharmaceutical Engineering,Shenyang pharmaceutical University,Shenyang 110015,China Shanghai Research center of biotechnology,Chinese Academy of Sciences,Shanghai 200233,China School of pharmaceutical Engineering Shenyang Pharmaceutical UniversityShenyang 110015 China Shanghai Research center of biotechnology Chinese Academy of SciencesShanghai 200233 China

A phage display single chain fragment variable library constructed on pⅢ protein of M13filamentous phage was screened using B-lymphocyte stimulator and FP248 as selective *** fourrounds of panning,there was a remarkable enrichment in the titer of bound *** phage cloneswere selected from the last round and screened by means of *** the antibody phages asprimary antibodies in Western blot,we developed a method for detecting the specific *** dilutions ofantibody phages depend on the affinity between antibody-displayed phage particles and antigens.

关键词： phage-display antibody Western blot

Expression of Recombinant Human FADD,Preparation of Its Polyclonal Antiserum and the Application in Immunoassays

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Cellular & Molecular Immunology 2008年第6期5卷 471-474页

作者： Faiz MMT Marikar The State Key Laboratory of pharmaceutical biotechnology and Jiangsu center of Hepatobiliary Diseases Nanjing University

The wild-type human Fas-associated death domain （FADD） protein was expressed as a His-tag fusion protein in Escherichia coll. Recombinant FADD proteins were purified under the denatured condition. After denatured protein purification, it was refolded and obtained at a yield of about 23 mg/L. Purified FADD exhibited as a homogenous band corresponding to the molecular weight of 31 kDa. Immunization of rabbits against the refolded FADD protein was allowed the production of high titre polyclonal antiserum. This new polyelonal antibody could recognize recombinant FADD protein in Western blot. Immunoreactivity was also observed in immunofiuorescence assay. The low cost polyclonal antiserum was applicable to extensive detection of FADD in various immunoassays. Cellular ＆ Molecular Immunology. 2008;5（6）：471-474.

关键词： FADD His-tag fusion protein polyclonal antibody immunofluorescence assay

Horizontal transfer of microRNAs: molecular mechanisms and clinical applications

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Protein & Cell 2012年第1期3卷 28-37页

作者： Xi Chen Hongwei Liang Junfeng Zhang Ke Zen Chen-Yu Zhang Jiangsu Engineering Research center for microRNA Biology and biotechnology State Key Laboratory of Pharmaceutical BiotechnologySchool of Life SciencesNanjing UniversityNanjing 210093China

A new class of RNA regulatory genes known as microRNAs(miRNAs)has been found to introduce a whole new layer of gene regulation in *** intensive studies of the past several years have demonstrated that miRNAs are not only found intracellularly,but are also detectable outside cells,including in various body fluids(***,plasma,saliva,urine and milk).This phenomenon raises questions about the biological function of such extracellular *** amounts of extracellular miRNAs are enclosed in small membranous vesicles(***,shedding vesicles and apoptotic bodies)or packaged with RNA-binding proteins(***-density lipoprotein,Argonaute 2 and nucleophosmin 1).These miRNAs may function as secreted signaling molecules to influence the recipient cell ***,secreted extracellular miRNAs may reflect molecular changes in the cells from which they are derived and can therefore potentially serve as diagnostic indicators of *** studies also point to the potential application of siRNA/miRNA delivery as a new therapeutic strategy for treating *** this review,we summarize what is known about the mechanism of miRNA *** addition,we describe the pathophysiological roles of secreted miRNAs and their clinical potential as diagnostic biomarkers and therapeutic *** believe that miRNA transfer between cells will have a significant impact on biological research in the coming years.

关键词： microRNA extracellular microRNA microRNA secretion horizontal transfer microvesicle exosome apoptotic body high-density lipoprotein Argonaute 2 nucleophosmin 1 diagnosis therapy

Identification of Ebola virus microRNAs and their putative pathological function

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Science China(Life Sciences) 2014年第10期57卷 973-981页

作者： LIANG HongWei ZHOU Zhen ZHANG SuYang ZEN Ke CHEN Xi ZHANG ChenYu Jiangsu Engineering Research center for microRNA Biology and biotechnology State Key Laboratory of Pharmaceutical BiotechnologySchool of Life Sciences Nanjing University

Ebola virus(EBOV),a member of the filovirus family,is an enveloped negative-sense RNA virus that causes lethal infections in humans and ***,more than 1000 people have been killed by the Ebola virus disease in Africa,yet no specific treatment or diagnostic tests for EBOV are *** this study,we identified two putative viral microRNA precursors(pre-miRNAs)and three putative mature microRNAs(miRNAs)derived from the EBOV *** production of the EBOV miRNAs was further validated in HEK293T cells transfected with a pcDNA6.2-GW/EmGFP-EBOV-pre-miRNA plasmid,indicating that EBOV miRNAs can be produced through the cellular miRNA processing *** also predicted the potential target genes of these EBOV miRNAs and their possible biological ***,this study reports for the first time that EBOV may produce miRNAs,which could serve as non-invasive biomarkers for the diagnosis and prognosis of EBOV infection and as therapeutic targets for Ebola viral infection treatment.

关键词： microRNA Ebola virus function

Nuclear microRNAs and their unconventional role in regulating non-coding RNAs

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Protein & Cell 2013年第5期4卷 325-330页

作者： Hongwei Liang Junfeng Zhang Ke Zen Chen-Yu Zhang Xi Chen Jiangsu Engineering Research center for microRNA Biology and biotechnology State Key Laboratory of Pharmaceutical BiotechnologySchool of Life SciencesNanjing UniversityNanjing 210093China

MicroRNAs (miRNAs) are small non-coding RNAs (ncRNAs) that are involved in post-transcriptional gene regulation. It has long been assumed that miRNAs exert their roles only in the cytoplasm, where they recognize their target protein-coding messenger RNAs (mRNAs), and result in translational repression or target mRNA degradation. Recent studies, however, have revealed that mature miRNAs can also be transported from the cytoplasm to the nucleus and that these nuclear miRNAs can function in an unconventional manner to regulate the biogenesis and functions of ncRNAs (including miRNAs and long ncRNAs), adding a new layer of complexity to our understanding of gene regulation. In this review, we summarize recent literature on the working model of these unconventional miRNAs and speculate on their biological significance. We have every reason to believe that these novel models of miRNA function will become a major research topic in gene regulation in eukaryotes.

关键词： microRNA nuclear microRNA non-coding RNA nucleus Argonaute Exportin Importin

Selective secretion of microRNA in CNS system

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Protein & Cell 2013年第4期4卷 243-247页

作者： Qipeng Zhang Jie Xu Qun Chen Xi Chen Ke Zen Chen-Yu Zhang Jiangsu Engineering Research center for microRNA Biology and biotechnology State Key Laboratory of Pharmaceutical BiotechnologySchool of Life SciencesNanjing UniversityNanjing 210093China

EXTRACELLULAR miRNAs ARE PRESENT IN VARIOUS BODY FLUIDS MicroRNAs(miRNAs)are a diverse class of endogenous small non-coding RNAs,which posttranscriptionally regulate gene expression by interacting with their binding sites in target ***,several studies have demonstrated that miRNAs are also detectable outside cells,and these miRNAs may be called extracellular *** miRNAs are found in various body fluids,including plasma and serum(Chen et al.,2008),saliva(Park et al.,2009),amniotic fl uid(Weber et al.,2010),Bronchial lavage(Molina-Pinelo et al.,2012;Weber et al.,2010),sputum(Xie et al.,2010),tears and urine(Weber et al.,2010),bile(Shigehara et al.,2011),seminal plasma(Wang et al.,2011),breast milk(Zhou et al.,2012),peritoneal fluid(Chen et al.,2012a).Finding the extracellular miRNAs extended the research field of miRNAs,and got a lot of fruitful research fi ndings.

关键词： al. Weber endogenous

Small RNA existed in commercial reverse transcriptase： primary evidence of functional small RNAs

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Protein & Cell 2015年第1期6卷 1-5页

作者： Jie Xu Xi Chen Donghai Li Qun Chen Zhen Zhou Dongxia Hou Jin Wang Qipeng Zhang Ke Zen Chen-Yu Zhang Jiangsu Engineering Research center for microRNA Biology and biotechnology State Key Laboratory of Pharmaceutical Biotechnology School of Life Sciences Nanjing University Nanjing 210093 China

SMALL RNA IN REVERSE TRANSCRIPTASE： TOO SMALL TO BE FOUND Significant background in no-template or no-primer RT-PCR is frequently noted. However, the underlying mechanism is still unclear. In this issue, Jie Xu and colleagues reveal that the com- mercial reverse transcriptases contain small RNAs. In AMV reverse transcriptase, they detected microRNAs derived from the virus host chick or other birds. This surprising finding at least partially explains the high background in some AMV reverse transcriptases based RT-PCR.

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