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Protective effects of anti-ricin A-chain RNA aptamer against ricin toxicity

World Journal of Gastroenterology 2008年第41期14卷 6360-6365页

作者： Shaoan Fan Feng Wu Frank Martiniuk Martha L Hale Andrew D Ellington kam-meng tchou-wong Department of Environmental Medicine New York University School of Medicine 57 Old Forge Road Tuxedo New York 10987 United States Department of Medicine New York University School of Medicine 550 First Avenue New York 10016 United States Integrated Toxicology Division U.S. Army Medical Research Institute of Infectious Diseases (USAMRIID) Fort Detrick Maryland 21702 United States Department of Chemistry and Biochemistry University of Texas at Austin Austin Texas 78712 United States

AIM： To investigate the therapeutic potential of an RNA ligand （aptamer） specific for the catalytic ricin A-chain （RTA）, the protective effects of a 31-nucleo- tide RNA aptamer （31RA）, which formed a high affinity complex with RTA, against ricin-induced toxicity in cell- based luciferase translation and cell cytotoxicity assays were evaluated. METHODS： To test the therapeutic potential of anti- RTA aptamers in Chinese hamster ovary （CliO） AA8 cells stably transfected with a tetracycline regulatable promoter, ricin ribotoxicity was measured us- ing luciferase and ricin-induced cytotoxicity was ascertained by MTS cell proliferation assay with tet- razolium compound [3-（4,5-dimethylthiazol-2-yl）- 5-（3-carboxymethoxyphenyl）-2-（4-sulfophenyl）-2H- tetrazolium]. RESULTS： Inhibition of protein synthesis by ricin in CliO AA8 cells resulted in diminished luciferase activity and treatment with polyclonal antibody against degly- cosylated RTA （dgA） neutralized the inhibitory effects of ricin on luciferase activity and protected against ricin-induced cytotoxicity as measured by MTS assay. The 31RA anti-RTA aptamer inhibited the translation of luciferase mRNA in cell-free reticulocyte translation assay. 31RA aptamer also partially neutralized the inhibitory effects of ricin on luciferase activity and partially protected against ricin-induced cytotoxicity in CliO AA8 cells. CONCLUSION： We have shown that anti-RTA RNA aptamer can protect against ricin ribotoxicity in cell- based luciferase and cell cytotoxicity assays. Hence, RNA aptamer that inhibits RTA enzymatic activity represents a novel class of nucleic acid inhibitor that has the potential to be developed as a therapeutic agent for the treatment of ricin intoxication.

关键词： Ricin inhibitor RNA aptamer Luciferase assay

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Correlation of N-myc downstream-regulated gene 1 expression with clinical outcomes of colorectal cancer patients of different race/ethnicity

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World Journal of Gastroenterology 2007年第20期13卷 2803-2810页

作者： Minori Koshiji Kensuke Kumamoto Keiichirou Morimura Yasufumi Utsumi Michiko Aizawa Masami Hoshino Shinji Ohki Seiichi Takenoshita Max Costa Thérèse Commes David Piquemal Curtis C Harris kam-meng tchou-wong Department of Environmental Medicine New York University School of Medicine 57 Old Forge Road Tuxedo NY 10987 United States Shinji Ohki Seiichi Takenoshita Second Department of Surgery Fukushima Medical University 1 Hikarigaoka Fukushima 960-1295 Japan Curtis Harris National Institutes of Health National Cancer Institute Building 37 Bethesda MD 20892 United States Department of Pathology Osaka City University Osaka Japan Department of Pathology Ohara General Hospital 6-11 Ohmachi Fukushima 960-8611 Japan Department of Surgery Ohara General Hospital 6-11 Ohmachi Fukushima 960-8611 Japan David Piquemal University Montpellier Ⅱ Montpellier France

AIM： To evaluate the role of N-myc downstream- regulated gene 1 （NDRG1） expression in prognosis and survival of colorectal cancer patients with different ethnic backgrounds. METHODS： Because NDRG1 is a downstream target of p53 and hypoxia inducible factor-1α （HIF-1α）, we examined NDRG1 expression together with p53 and HIF-1α by irnmunohistochernistry. A total of 157 colorectal cancer specimens including 80 from Japanese patients and 77 from US patients were examined. The correlation between protein expression with clinicopathological features and survival after surgery was analyzed. RESULTS： NDRG1 protein was significantly increased in colorectal tumor compared with normal epithelium in both Japanese and US patient groups. Expression of NDRG1 protein was significantly correlated with lymphatic invasion, venous invasion, depth of invasion, histopathological type, and Dukes＇ stage in Japanese colorectal cancer patients. NDRG1 expression was correlated to histopathological type, Dukes＇ stage and HIF-1α expression in US-Caucasian patients but not in US-African American patients. Interestingly, Kaplan-Meier survival analysis demonstrated that NDRG1 expression correlated significantly with poorer survival in US-African American patients but not in other patient groups. However, in p53-positive US cases, NDRG1 positivity correlated significantly with better survival. In addition, NDRG1 expression also correlated significantly with improved survival in US patients with stages Ⅲ and IV tumors without chemotherapy. In Japanese patients with stages Ⅱ and Ⅲ tumors, strong NDRG1 staining in p53- positive tumors correlated significantly with improved survival but negatively in patients without chemotherapy. CONCLUSION： NDRG1 expression was correlated with various clinicopathological features and clinical outcomes in colorectal cancer depending on the race/ethnicity of the patients. NDRG1 may serve as a biological basis for the disparity of clinical outcomes of colorectal canc

关键词： NDRG1 expression Colorectal cancer Race Ethnicity Clinical outcomes

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Protective effects of anti-ricin A-chain antibodies delivered intracellularly against ricin-induced cytotoxicity

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World Journal of Biological Chemistry 2010年第5期1卷 188-195页

作者： Frank Martiniuk Seth Pincus Sybille Müller Heinz Kohler kam-meng tchou-wong Department of Medicine New York University School of Medicine550 First AvenueNew YorkNY 10016 United States Department of Research Children's Hospital200 Henry Clay AvenueNew OrleansLA 70118United States Immpheron Inc.5235 Athens-Boonesboro RoadLexingtonKY 40509United States Markey Cancer Center University of Kentucky800 Rose StreetLexingtonKY 40536-0096United States Department of Environmental Medicine New York University School of Medicine57 Old Forge RoadTuxedoNY 10987United States

AIM:To evaluate the ability of anti-ricin A-chain antibodies,delivered intracellularly,to protect against ricininduced cytotoxicity in RAW264.7 cells. METHODS:Anti-deglycosylated ricin A-chain antibody and RAC18 anti-ricin A-chain monoclonal antibody were delivered intracellularly by encapsulating in liposomes or via conjugation with the cell-penetrating MTS-transport ***264.7 cells were incubatedwith these antibodies either before or after ricin *** changes in cytotoxicity were estimated by MTT ***-localization of internalized antibody and ricin was evaluated by fluorescence microscopy. RESULTS:Internalized antibodies significantly increased cell viability either before or after ricin exposure compared to the unconjugated *** microscopy confirmed the co-localization of internalized antibodies and ricin inside the cells. CONCLUSION:Intracellular delivery of antibodies to neutralize the ricin toxin after cellular uptake supports the potential use of cell-permeable antibodies for postexposure treatment of ricin intoxication.

关键词： Ricin inhibitors Antibody internalization Cell-penetrating peptide Cell-permeable antibodies Liposome-encapsulated antibodies

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d-limonene prevents ultraviolet irradiation:Induced cyclobutane pyrimidine dimers in Skh1 mouse skin

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World Journal of Dermatology 2014年第3期3卷 64-72页

作者： Ahmed N Uddin Feng Wu Ivica Labuda kam-meng tchou-wong Fredric J Burns Department of Environmental Medicine New York University School of Medicine Tuxedo NY 10987 United States

AIM: To establish whether d-limonene can protect against induction of cyclobutane pyrimidine dimers(CPDs) and sunburn in ultraviolet irradiation(UVR) irradiated mouse skin. METHODS: The d-limonene was given in 4 daily oral 20 μL aliquots at different concentrations as follows: 100%, 10% or 1% in liponate and 100% liponate as control. One day after the final d-limonene treatment, the mice were anesthetized with i.p. sodium pentobarbital and placed in boxes to allow a rectangular(2 cm × 4 cm) region of dorsal skin to be irradiated with a single, ultraviolet radiation dose of 1.5 kJ /m2. Skin samples from UVR irradiated area were obtained at 5 min after UVR exposure for CPD detection, at 6 d after UVR exposure, skin samples were obtained for in situ analysis for N-myc downstream regulating gene 1(NDRG1)(a stress response gene), proliferating cell nuclear antigen(PCNA)(an S-phase marker) and filaggrin(a barrier integrity gene). Based on immunohistochemistry staining, the number of CPD, NDRG1 and PCNA positive cells, as well as unstained cells was counted in 3 different individually selected areas and percentage of positive cells was established. RESULTS: CPD reduction occurred as follows: liponate only-none; 1% d-limonene-54.3% reduction of CPDs; 10% d-limonene-73.4% reduction of CPDs; 100% d-limonene-86.1% reduction of CPDs, the latter equivalent to a UV dose of only 0.21 k J/m2. Sunburn was also dose-dependently reduced by d-limonene. The NDRG1 protein was strongly induced by UVR(70.0% ± 10.4% positive cells), but 1% d-limonene reduced the response to 64.6% ± 9.2%, 10% d-limonene reduced the response to 16.2% ± 3.4% and 100% d-limonene reduced the response to 6.3% ± 1.7%. Similarly, PCNA was 52.4% ± 9.9% positive in UVR exposed skin, and 1% d-limonene reduced it to 42.9% ± 8.1%, 10% d-limonene reduced it to 36.2% ± 6.7% and 100% d-limonene reduce it to 13.8% ± 3.4%. NDRG1 and PCNA were increased by d-limonene or UVR separately, but combined they produced less than eit

关键词： Sunburn Ultraviolet irradiation d-limonene Cyclobutane pyrimidine dimers N-myc downstream regulating gene 1 Proliferating cell nuclear antigen

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A converged ubiquitin-proteasome pathway for the degradation of TOC and TOM tail-anchored receptors

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Journal of Integrative Plant Biology 2024年第5期66卷 1007-1023页

作者： Meijing Yang Shuai Chen Shey-Li Lim Lang Yang Jia Yi Zhong Koon Chuen Chan Zhizhu Zhao kam-Bo wong Junqi Wang Boon Leong Lim School of Biological Sciences University of Hong KongPokfulam 999077Hong KongChina Department of Biology Southern University of Science and TechnologyShenzhen 518055China School of Biomedical Engineering Shenzhen UniversityShenzhen 518060China School of Life Sciences The Chinese Universityof Hong KongShatin 999077HongKongChina State Key Laboratory of Agrobiotechnology The Chinese University of Hong KongShatin 999077Hong KongChina HKU Shenzhen Institute of Research and Innovation Shenzhen 518052China

In plants,thousands of nucleus-encoded proteins translated in the cytosol are sorted to chloroplasts and mitochondria by binding to specific receptors of the TOC(translocon on the outer chloroplast membrane)and the TOM(translocon on the outer mitochondrial membrane)complexes for import into those *** degradation pathways for these receptors are ***,we discovered a converged ubiquitin-proteasome pathway for the degradation of Arabidopsis thaliana TOC and TOM tail-anchored *** receptors are ubiquitinated by E3 ligase(s)and pulled from the outer membranes by the AAA+adenosine triphosphatase CDC48,after which a previously uncharacterized cytosolic protein,transmembrane domain(TMD)-binding protein for tail-anchored outer membrane proteins(TTOP),binds to the exposed TMDs at the C termini of the receptors and CDC48,and delivers these complexes to the 26S proteasome.

关键词： 26S proteasome CDC48 SP1 TOC TOM ubiquitination

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Computational Approaches for Traditional Chinese Painting: From the “Six Principles of Painting” Perspective

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Journal of Computer Science & Technology 2024年第2期39卷 269-285页

作者：张玮张建伟黄锦畦王懿芳封颖超杰王璐玮陈为 State Key Laboratory of CAD&CG Zhejiang UniversityHangzhou 310058China Department of Computer Science and Engineering The Hong Kong University of Science and Technology Hong Kong 999077China Kellogg School of Management Northwestern UniversityEvanston 60208U.S.A Laboratory of Art and Archaeology Image Zhejiang UniversityHangzhou 310058China

Traditional Chinese painting(TCP)is an invaluable cultural heritage resource and a unique visual art *** recent years,there has been a growing emphasis on the digitalization of TCP for cultural preservation and *** resulting digital copies have enabled the advancement of computational methods for a structured and systematic understanding of *** explore this topic,we conduct an in-depth analysis of 94 pieces of *** examine the current use of computer technologies on TCP from three perspectives,based on numerous conversations with ***,in light of the“Six Principles of Painting”theory,we categorize the articles according to their research focus on artistic ***,we create a four-stage framework to illustrate the purposes of TCP ***,we summarize the popular computational techniques applied to *** work also provides insights into potential applications and prospects,with professional opinion.

关键词： traditional Chinese painting(TCP) digital humanities cultural heritage computer vision deep learning

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Effects of nitric oxide on gastric ulceration induced by nicotine and cold-restraint stress

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World Journal of Gastroenterology 2004年第4期10卷 594-597页

作者： Bo-ShengQui Qi-BingMei LiLiu kam-meng tchou-wong DepartmentsofMedicine EnvironmentalMedicineandMicrobiologyNewYorkUniversitySchoolofMedicineNewYorkUSA DepartmentofPharmacology TheFourthMilitaryMedicalUniversityXianChina

AIM:Stress induces gastric ulceration in human and experimental animals. People tend to smoke more cigarettes when under stress. Nitric oxide (NO) and nicotine have opposing effects on gastric integrity. The present study examined the possible therapeutic benefit of NO in nicotine-treated rats with stress-induced gastric ***:Rats drank a nicotine solution while control rats drank tap water for 20 days. The alkoloid was then replaced by water with or without supplementation of isosorbide dinitrate (NO donor) for an additional 10 days. Isosorbide dinitrate was given twice shortly before experiments (acute) or three times daily by oral gavages for 10 days after the rats stopped drinking nicotine solution. At the end of experiments,ulcer index, gastric adhesion mucus content and MPO activity were measured and ***:Nicotine treatment decreased gastric mucus content and intensified stress-induced gastric ulcer. A higher ulcer index persisted even after the rats stopped drinking nicotine solution for 10 days. Acute NO donor showed no benefit on both mucus and ulcer index in nicotine treatment or/and stress *** NO donor treatment reversed the worsening action of nicotine in stomach. Stress increased gastric mucosal myeloperoxidase (MPO) activity, which was antagonized by chronic NO treatment. However, nicotine was unlikely to change mucosal MPO ***:The intensifying action of nicotine on stressinduced gastric ulceration persists for 10 days after *** treatment significantly decreases gastric mucus content that can be restored by chronic NO donor *** present study suggests that NO antagonizes the ulcerogenic action of nicotine through a cytoprotective way.

关键词：烟碱胃溃疡一氧化氮动物模型压力病理机制

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铜绿假单胞菌对肺血管内皮细胞功能的影响及其机制

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中国医学科学院学报 2024年第1期46卷 1-10页

作者：张雷王俊轶何翔吴敏李国平黄锦伟澳门科技大学中药质量研究国家重点实验室中国澳门999078 成都市第三人民医院呼吸健康研究所过敏与精准医学实验室成都610000 北达科他大学医学与健康科学院生物医学科学系美国北达科他州大福克斯58203 中国科学院大学温州研究院浙江温州325000

目的探究铜绿假单胞菌(PA)感染对肺血管内皮细胞功能的影响,并探讨其加重小鼠肺部炎症反应的可能机制。方法PAO1菌株感染人肺微血管内皮细胞HULEC-5a 2 h后,采用逆转录实时荧光定量PCR(RT-qPCR)检测自噬相关基因5(ATG5)、6-磷酸果糖-2-... 详细信息

目的探究铜绿假单胞菌(PA)感染对肺血管内皮细胞功能的影响,并探讨其加重小鼠肺部炎症反应的可能机制。方法PAO1菌株感染人肺微血管内皮细胞HULEC-5a 2 h后,采用逆转录实时荧光定量PCR(RT-qPCR)检测自噬相关基因5(ATG5)、6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶3(PFKFB3)和钙黏蛋白5(CDH5)的mRNA表达,免疫荧光检测ATG5、PFKFB3和血管内皮钙黏蛋白(VE-cadherin)的蛋白表达。采用小干扰RNA(siRNA)分别敲减ATG5、PFKFB3后,RT-qPCR检测ATG5、PFKFB3和CDH5的mRNA表达,免疫荧光检测PFKFB3和VE-cadherin的蛋白表达,并采用乳酸测定试剂盒检测细胞内乳酸的水平。采用PAO1菌株感染小鼠后,肺组织切片病理染色观察小鼠肺部炎症,激光共聚焦显微镜观察并分析荧光标记的内皮细胞PFKFB3和VE-cadherin的表达量。结果与对照组比较,PAO1感染HULEC-5a细胞后,PFKFB3 mRNA和蛋白表达均增加(P均<0.05),CDH5 mRNA表达减少(P=0.023),VE-cadherin蛋白表达减少(P<0.001),且连续性被破坏,乳酸含量升高(P=0.017)。与PAO1感染HULEC-5a细胞比较,PAO1感染敲减PFKFB3的HULEC-5a细胞后,CDH5 mRNA表达增加(P=0.043),VE-cadherin荧光连续性得到恢复,乳酸含量降低(P=0.047);PAO1感染敲减ATG5的HULEC-5a细胞后,PFKFB3 mRNA(P=0.013)和蛋白表达(P=0.003)均增加,CDH5 mRNA(P=0.020)和VE-cadherin蛋白表达(P=0.001)均减少,乳酸含量升高(P=0.015)。PAO1感染小鼠后,病理HE染色结果显示,肺泡内明显的红细胞渗漏,炎症细胞浸润,肺泡间隔增宽和部分血管内皮细胞脱落。免疫荧光染色结果显示,与正常小鼠比较,PAO1感染小鼠内皮细胞PFKFB3表达增加,VE-cadherin荧光减弱,且不连续。结论PA可能通过AGT5调控PFKFB3通路,破坏肺血管内皮细胞功能,进而加重小鼠肺部炎症反应。

关键词：铜绿假单胞菌肺血管内皮细胞 6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶3 自噬相关基因5 感染

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Schematic interface of sound creation for computer animators

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Journal of Zhejiang University-Science A(Applied Physics & Engineering) 2006年第7期7卷 1141-1151页

作者： TONG kam-pang Maya wong kam-wah School of Creative Media City University of Hong Kong Hong Kong China

As an audiovisual medium, computer animations require superior image quality and professional soundtrack to lead audiences into their fascinating virtual world. Without sound, the impact of storytelling is reduced or the story is even not under- standable. Despite the importance of sound, most animators are unfamiliar with sound editing software. Limited budget projects such as independent or student works have difficulty hiring sound professionals to create tailor-made soundtrack. Therefore, we need a suitable sound tool to express their individual ideas. In this paper, we propose an approach using schematic for both computer animation and sound. Our approach provides (1) physical simulation on sound through animation parameters and (2) a new channel for animators to add aesthetic values in sound through their experience of using schematics in existing animation software. We demonstrate our idea through several examples, such as Doppler shift, obstacle effect, importance, energetic, and sputtering effect.

关键词： Computer animation Multimedia Digital media Schematics Physical simulation Aesthetic Sound effects Soundtrack

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Development of An ICR Mouse Bioassay for Toxicity Evaluation in Neurotoxic Poisoning Toxins-Contaminated Shellfish

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Biomedical and Environmental Sciences 2013年第5期26卷 346-364页

作者： wong Chun Kwan HUNG Patricia kam Kai Man Biotoxin Laboratory Microbiology DivisionPublic Health Laboratory Services BranchCentre for Health ProtectionDepartment of HealthShek Kip MeiKowloon

Objective To develop an ICR （female） mouse bioassay （MBA） for toxicity confirmation and evaluation of neurotoxins （brevetoxins）-contaminated shellfish. Methods Brevetoxins （BTX-B） as a causative agent of neurotoxic shellfish poisoning （NSP） under different shellfish matrices were intraperitoneally injected at different doses into mice to study their toxic effects and to differentiate the range of lethal and sublethal dosages. Their sensitivity and specificity were analyzed with 2 competitive ELISA kits for quantitative determination of standard BTX-B and dihydroBTX-B under different shellfish matrix-diluent combinations. Detection rates of MBA and two antibody-based assays for BTX-B from field NSP-positive shellfish samples were compared. Results BTX-B could be detected in shellfish tissues at concentration of 50-400 μg/100 g under shellfish matrix-Tween-saline media, which were appropriate to identify toxic shellfish at or above the regulatory limit （80 μg/100 g shellfish tissues）. The LD 50 identified was 455 g/kg for BTX-B under general shellfish matrices （excluding oyster matrices） dissolved in Tween-saline. The presence of shellfish matrices, of oyster matrices in particular, retarded the occurrence of death and toxicity presentation in mice. Two antibody-based assays, even in the presence of different shellfish matrix-diluent combinations, showed acceptable results in quantifying BTX-B and dihydroBTX-B well below the regulatory limit. Conclusion The two ELISA analyses agree favorably （correlation coefficient, r 0.96; Student＇s t-tests, P〉0.05） with the developed bioassay.

关键词： Antibody-based assay Brevetoxin ICR female mice LD 50 Mouse bioassay Neurotoxic shellfish poisoning

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