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中华耳鼻咽喉头颈外科杂志 2016年第1期51卷 43-49页

作者：蔺林戴飞孙庭钰复旦大学附属华山医院耳鼻咽喉头颈外科上海200040

目的研究Orail抗体腹腔内注射是否可以改善小鼠变应性鼻炎（allergicrhinitis，AR）的炎症状态。方法选取健康雌性BALB／c小鼠（SPF级）24只随机数字表法分为4组（AR组、对照组、实验组1、实验组2），每组6只。建立AR小鼠模型（对照组... 详细信息

目的研究Orail抗体腹腔内注射是否可以改善小鼠变应性鼻炎（allergicrhinitis，AR）的炎症状态。方法选取健康雌性BALB／c小鼠（SPF级）24只随机数字表法分为4组（AR组、对照组、实验组1、实验组2），每组6只。建立AR小鼠模型（对照组以磷酸盐缓冲液建模），以不同剂量Orail抗体（实验组1、2分别为100、150μg）行腹腔内注射，观察小鼠在Orail抗体干预后打喷嚏和挠鼻次数的改变，检测小鼠鼻黏膜内嗜酸粒细胞浸润数的变化。酶联免疫吸附实验（enzymelinkedimmunosorbentassay，ELISA）和实时反转录聚合酶链反应（real—timereversetranscription—polymerasechainreaction，real—timeRT—PcR）分别检测小鼠鼻黏膜中Orail浓度、鼻腔灌洗液（nasallavagefluid，NLF）中组织胺、嗜酸粒细胞阳离子蛋白（eosionphilcationprotein，EcP）和白细胞介素（interlukin，IL）1B、4、5、6的浓度以及外周血Th2细胞中Orail、血清中IL-d、IL-5的浓度，并分别测试小鼠鼻黏膜和外周血Th2细胞内上述物质的mRNA含量。以GraphPadPrism5统计软件进行数据分析。结果Orail抗体干预后小鼠打喷嚏、挠鼻次数和鼻黏膜内嗜酸粒细胞浸润数较AR组差异有统计学意义（t100g值分别为7．88、9．92、4．30，t150μg值分别为16．43、16．31、9．35，P值均〈0．01）；小鼠鼻黏膜中Orail、NLF中组织胺、EcP和IL-1p、IL-4、IL-5、IL-6的浓度分别为（0．186±0．015）μg／ml、（6．618±0．180）ng／ml、（2．555±0．031）ng／ml、（85．26±2．94）pc,／ml、（55．12±1．21）pg／ml、（58．454-2．11）pc,／ml、（77．124-2．13）pg／ml（实验组1），（0．089±0．003）μg／ml、（4．501±0．310）ng／ml、（1．260±0．017）ng／ml、（48．49±2．12）pg／ml、（33．15±0．87）pg／ml、（38．24±0．95）pg／ml、（51．72±0．81）pg/ml、（实验组2），均较AR组差异有统计学意义（t值分别为3．29、10．44、9．45、17．53、74．53、87．06、3．98；8．54、13．32、23．00、20．89、80．73、103．70、13．34，P值均〈0．01）；鼻黏膜和NLF中相应mRNA的含量亦呈现相同变化规律。而其外周血Th2细胞中Orail、血清中IL-4和IL-5的浓度及Th2细胞内上述物质mRNA的含量差异无统计学意义（P值均〉0．05）；且100μgOrail抗体的治疗作用明显低于150μg的治疗作用，差异有统计学意义（P值均〈0．05）。结论Orail抗体腹腔内注射可以改善小鼠AR的症状和炎症程度，Orail有可能成为治疗AR新的研究方向。

关键词：鼻炎变应性常年性疾病模型动物小鼠近交BALB c 抗体治疗应用

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ERK/cREB信号通路在异菝葜皂苷元促进Aβ损伤SH-SY5Y细胞BDNF表达中的作用

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上海交通大学学报（医学版） 2016年第5期36卷 642-647页

作者：张瑞夏志明孙小余李加梅张永芳胡雅儿上海交通大学医学院细胞调控研究室上海200025 上海交通大学医学院转化医学协同创新中心上海200025

目的探讨细胞外信号调节激酶/c AMP反应元件结合蛋白(ERK/cREB)信号通路在异菝葜皂苷元(SMI)促进β-淀粉样肽(Aβ)损伤SH-SY5Y细胞脑源性神经营养因子(BDNF)表达中的作用及其机制。方法建立Aβ损伤SH-SY5Y细胞模型,采用RT-PcR法检测BDNF... 详细信息

目的探讨细胞外信号调节激酶/c AMP反应元件结合蛋白(ERK/cREB)信号通路在异菝葜皂苷元(SMI)促进β-淀粉样肽(Aβ)损伤SH-SY5Y细胞脑源性神经营养因子(BDNF)表达中的作用及其机制。方法建立Aβ损伤SH-SY5Y细胞模型,采用RT-PcR法检测BDNF m RNA的表达,用Western blotting的方法检测p cREB和p ERK的变化,最后通过阻断实验确认ERK/cREB信号通路在SMI促进BDNF m RNA表达中的作用。结果 SMI能促进Aβ损伤SH-SY5Y细胞BDNF m RNA的表达(P=0.000),并能上调信号分子cREB和ERK1/2的磷酸化水平(P=0.001,P=0.000)。用RNA干扰的方法阻断cREB的表达后,SMI促进BDNF m RNA表达的作用完全消失;用U0126阻断ERK1/2磷酸化后,SMI促进p cREB水平升高的作用完全消失。结论 SMI通过ERK/cREB通路促进Aβ损伤SH-SY5Y细胞BDNF m RNA的表达。

关键词：异菝葜皂苷元阿尔茨海默症脑源性神经营养因子 c AMP反应元件结合蛋白细胞外信号调节激酶1/2

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中华大蟾蜍色氨酸羟化酶BbgTPH1基因的克隆、原核表达及功能鉴定

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药学学报 2016年第7期51卷 1158-1164页

作者：刘彦芳陈曦凌鸿刘东春夏明钰王东沈阳药科大学中药学院辽宁沈阳110016 沈阳药科大学中韩分子生药学研究室辽宁沈阳110016 沈阳药科大学生命科学与生物制药学院辽宁沈阳110016

色氨酸羟化酶(tryptophan hydroxylase I,TPH1)是催化色氨酸生成5-羟色胺的限速酶,而5-羟色胺不仅是蟾酥的主要活性成分,同时也是中枢神经系统重要的神经递质,具有重要的生理功能。本研究从中华大蟾蜍耳后腺文库中克隆得到中华大蟾蜍色... 详细信息

色氨酸羟化酶(tryptophan hydroxylase I,TPH1)是催化色氨酸生成5-羟色胺的限速酶,而5-羟色胺不仅是蟾酥的主要活性成分,同时也是中枢神经系统重要的神经递质,具有重要的生理功能。本研究从中华大蟾蜍耳后腺文库中克隆得到中华大蟾蜍色氨酸羟化酶c DNA。构建原核重组融合表达质粒p MAL-Bbg TPH1,将其导入Escherichia coli TB1中,用IPTG诱导重组菌并优化诱导表达条件。以色氨酸为底物进行体外酶促反应,采用Lc-MS法鉴定反应产物。结果显示Bbg TPH1基因c DNA全长为1 984 bp,开放阅读框1 443 bp,编码480个氨基酸,推测蛋白质分子质量为55.2 k Da,理论等电点为5.58,具有芳香族类羟化酶超基因家族的保守结构域和芳香族类羟化酶家族特有的特征信号序列。氨基酸比对分析表明,该基因编码的氨基酸与其他物种TPH1蛋白序列具有较高的同源性。系统进化树显示Bbg TPH1与非洲爪蟾的TPH1亲缘关系最近。Bbg TPH1在*** TB1中能够高效表达,其优化表达条件为诱导温度20℃、IPTG浓度0.5 mmol·L-1。酶活检测结果显示,重组Bbg TPH1具有较高催化活性,可将色氨酸转化为5-羟色氨酸。本研究首次获得了蟾蜍属动物色氨酸羟化酶全长c DNA序列,并验证了其催化功能,为进一步研究蟾蜍中蟾毒色胺类化合物的生物合成分子机制以及次生代谢调控等提供了科学依据。

关键词：中华大蟾蜍色氨酸羟化酶 c DNA 原核表达

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comparative Analysis of Rigid PVc Foam Reinforced with class c and class F Fly Ash

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Journal of Minerals and Materials characterization and Engineering 2014年第6期2卷 554-565页

作者： Parisa Khoshnoud Subhashini Gunashekar Murtatha M. Jamel Nidal Abu-Zahra Materials Science and Engineering Department University of Wisconsin-Milwaukee Milwaukee USA

Fly ash particles are usually spherical and based on their chemical composition;they are categorized into two classes: c and F. This study compares the microstructural, mechanical and thermal properties of extruded rigid PVc foam composites reinforced with class c and class F fly ash. The mechanical properties: such as tensile and flexural strength of composites containing class c fly ash were superior to the composites containing class F fly ash particles. composites containing 6 phr class c fly ash showed a 24% improvement in the tensile strength in comparison to a mere 0.5% increase in composites reinforced with class F fly ash. Similarly, the addition of 6 phr of class F fly ash to the PVc foam matrix resulted in a 5.74% decrease in the flexural strength, while incorporating the same amount of class c fly ash led to a 95% increase in flexural strength. The impact strength of the composites decreased as the amount of either type of fly ash increased in the composites indicating that fly ash particles improve the rigidity of the PVc foam composites. No significant changes were observed in the thermal properties of the composites containing either type of fly ash particles. However, the thermo-mechanical properties measured by DMA indicated a steep increase in the viscoelastic properties of composites reinforced with class c flyash. The microstructural properties studied by Scanning Electron Microscopy (SEM) confirmed that fly ash particles were mechanically interlocked in the PVc matrix with good interfacial interaction with the matrix. However, particle agglomeration and debonding was observed in composites reinforced with higher amounts of fly ash.

关键词： Rigid PVc Foam class c Fly Ash class F Fly Ash Thermo-Mechanical Properties Morphology

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Anti-rods/rings autoantibody generation in hepatitis cpatients during interferon-α/ribavirin therapy

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World Journal of Gastroenterology 2016年第6期22卷 1966-1974页

作者： Gerson Dierley Keppeke S John calise Edward KL chan Luis Eduardo c Andrade Rheumatology Division Escola Paulista de MedicinaUniversidade Federal de São PauloSP 04023-062Brazil Department of Oral Biology University of FloridaGainesvilleFL 32610-0424United States 3 Immunology DivisionFleury Medicine and Health Laboratories São PauloSP 04102-050Brazil

chronic inflammation associated with hepatitis c virus(HcV) infection can lead to disabling liver diseases with progression to liver cirrhosis and hepatocellular carcinoma. Despite the recent availability of more effective and less toxic therapeutic options, in most parts of the world the standard treatment consists of a weekly injection of pegylated interferon α(IFN-α) together with a daily dose of ribavirin. HcV patients frequently present circulating non-organ-specific autoantibodies demonstrating a variety of staining patterns in the indirect immunofluorescence assay for antinuclear antibodies(ANA). Between 20% to 40% of HcV patients treated with IFN-α and ribavirin develop autoantibodies showing a peculiar ANA pattern characterized as rods and rings(RR) structures. The aim of this article is to review the recent reports regarding RR structures and anti-rods/rings(antiRR) autoantibody production by HcV patients after IFN-α/ribavirin treatment. Anti-RR autoantibodies first appear around the sixth month of treatment and reach a plateau around the twelfth month. After treatment completion, anti-RR titers decrease/disappear in half the patients and remain steady in the other half. Some studies have observed a higher frequency of anti-RR antibodies in relapsers, i.e., patients in which circulating virus reappears after initially successful therapy. The main target of anti-RR autoantibodies in HcV patients is inosine-5'-monophosphate dehydrogenase 2(IMPDH2), the rate-limiting enzyme involved in the guanosine triphosphate biosynthesis pathway. Ribavirin is a direct IMPDH2 inhibitor and is able to induce the formation of RR structures in vitro and in vivo. In conclusion, these observations led to the hypothesis that anti-RR autoantibody production is a human model of immunologic tolerance breakdown that allows us to explore the humoral autoimmune response from the beginning of the putative triggering event: exposure to ribavirin and interferon.

关键词： Rods and rings autoantibodies Hepatitis c ribavirin Interferon-α

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Significance of hepatitis virus infection in the oncogenicinitiation of hepatocellular carcinoma

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World Journal of Gastroenterology 2016年第4期22卷 1497-1512页

作者： caecilia Hc Sukowati Korri E El-Khobar Susan I Ie Beatrice Anfuso David H Muljono claudio Tiribelli Fondazione Italiana Fegato ONLUS AREA Science Park Basovizza34149 TriesteItaly Department of Medicine Surgeryand Health SciencesUniversity of TriesteTeaching Hospital Cattinara34149 TriesteItaly Laboratory of Hepatitis and Emerging Disease Eijkman Institute for Molecular BiologyJakarta 10430Indonesia Storr Liver Centre The Westmead Institute for Medical ResearchThe University of Sydney at Westmead HospitalWestmead NSW 2145Australia

Hepatocellular carcinoma(Hcc) is one of the most common causes of cancer-related death worldwide. chronic infection of hepatitis B virus(HBV) and/or hepatitis c virus(HcV) is a major risk factor in the development of the Hcc, independently from excessive alcohol abuse and metabolic disease. Since the biology of HBV and HcV is different, their oncogenic effect may go through different mechanisms, direct and/or indirect. Viral hepatitis infection is associated with cellular inflammation, oxidative stress, DNA damage, that may lead to subsequent hepatic injuries such as chronic hepatitis, fibrosis, cirrhosis, and finally Hcc. Direct oncogenic properties of these viruses are related with their genotypic characteristics and the ability of viral proteins to interact with host proteins, thus altering the molecular pathways balance of the cells. In addition, the integration of HBV DNA, especially the gene S and X, in a particular site of the host genome can disrupt chromosomal stability and may activate various oncogenic mechanisms, including those in hematopoietic cells. Recently, several studies also had demonstrated that viral hepatitis could trigger the population of hepatic cancer stem cells. This review summarize available pre-clinical and clinical data in literature regarding oncogenic properties of HBV and HcV in the early initiation of Hcc.

关键词： Hepatocellular carcinoma Hepatitis B virus Hepatitis c virus Oncogenicity Viral pathogenicity

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Evaluation of Thermocyclic Oxidation Behavior of HVOF Sprayed (cr 3 c 2 -35% Nicr) + 5% Si coatings on Boiler Tube Steels

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Procedia Materials Science 2014年 5卷 398-407页

作者： B. Somasundaram Ravikiran Kadoli M.R. Ramesh Department of Mechanical Engineering NITK Surathkal Srinivasanagar 575025 India

In the present study, the alloying element Si is blended with (cr 3 c 2 -35% Nicr) powder and this composite powder is HVOF sprayed on three different Fe-based steels namely SA213-T22, MDN-310 and SUPERFER 800H. Oxidation studies have been conducted on the coated as well as uncoated specimens at 700 o c in air under cyclic conditions. The weight change method was used to establish the kinetics of oxidation. (cr 3 c 2 -35% Nicr) + 5% Si coated steels showed slow oxidation kinetics and considerably lower weight gains than that of uncoated steels. The superior performance of (cr 3 c 2 -35% Nicr) + 5% Si coatings can be attributed due to the formation of a compact and protective oxide layer consisting of cr 2 O 3 , SiO 2 , Nicr 2 O 4 and Ni 2 (SiO 4 ) phases. The uncoated T22 steel suffered a catastrophic degradation in the form of intense spalling of the scale. The combined technique of X-ray diffraction (XRD), scanning electron microscopy/energy dispersive X-ray analysis (***) and X-ray mapping were used to analyze the oxidized products of coated and bare substrate steels.

关键词： Protective coatings HVOF Thermal spray process High temperature Oxidation cr 3 c 2 -Nicr Alloying element SEM

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弓状核内胱硫醚β-合成酶通过PKc的上调和GluN2B磷酸化参与慢性胰腺炎大鼠腹部痛觉过敏（英文）

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生理学报 2016年第5期68卷 575-584页

作者：郑航朱宏岩张晓宇王蒙肖颖徐广银蒋星红苏州市疼痛基础研究和临床治疗重点实验室苏州大学神经生物学和生理学系神经科学研究所苏州215123

有报道显示初级感觉神经元中的硫化氢(H2S)参与内脏痛敏的形成,但其在中枢神经系统中的作用却鲜为人知。该研究旨在探讨下丘脑弓状核(arcuate nucleus,ARc)内H2S和其内源性合成酶是否参与慢性胰腺炎(chronic pancreatitis,cP)腹部痛觉... 详细信息

有报道显示初级感觉神经元中的硫化氢(H2S)参与内脏痛敏的形成,但其在中枢神经系统中的作用却鲜为人知。该研究旨在探讨下丘脑弓状核(arcuate nucleus,ARc)内H2S和其内源性合成酶是否参与慢性胰腺炎(chronic pancreatitis,cP)腹部痛觉过敏及其潜在机制。成年雄性Sprague-Dawley大鼠胰管内注射三硝基苯磺酸(trinitrobenzene sulfonic acid,TNBS)诱导产生cP模型,运用von Frey filament(VFF)评测大鼠腹部对机械刺激的反应频率,运用Western blot检测ARc内蛋白表达水平。TNBS注射4周后ARc内胱硫醚-β-合成酶(cystathionineβ-synthetase,cBS)表达显著上调,而胱硫醚-γ-裂解酶(cystathionine-γ-lyase,cSE)的表达没有明显改变;cP显著提高了NMDA受体Glu N2B亚单位的磷酸化水平,而总Glu N2B不变;cP也显著上调了蛋白激酶cγ(PKcγ)在ARc的表达。ARc内微量注射cBS的抑制剂O-(羧甲基)羟胺半盐酸盐(AOAA)能显著降低cP大鼠的腹部疼痛,也翻转了cP大鼠ARc中p-Glu N2B和PKcγ的上调;ARc内微量注射Glu N2B抑制剂或PKc特异性抑制剂白屈菜赤碱能显著减轻cP大鼠的腹部痛觉过敏,PKc特异性抑制剂能减少p-Glu N2B表达。总之,以上数据表明,cP引起了ARc内cBS的上调,并可能通过PKcγ介导了Glu N2B的活化,参与cP痛觉过敏。本研究在一定程度上揭示了cP痛觉过敏产生的中枢机制,并有助于发现cP疼痛治疗的新靶标。

关键词：弓状核慢性胰腺炎腹部痛觉过敏硫化氢 NMDA 受体蛋白激酶 c

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阿魏酸对脂多糖损伤的Pc12细胞和大鼠海马神经元细胞的保护作用

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中国药理学与毒理学杂志 2016年第4期30卷 330-337页

作者：黄浩马增春王宇光高月北京工业大学生命科学与生物工程学院北京100124 军事医学科学院放射与辐射医学研究所北京100850

目的探讨阿魏酸（FA）对脂多糖（LPS）诱导的Pc12细胞和大鼠海马神经元损伤的保护作用,并探讨其可能的作用机制。方法体外实验：FA 2.5～40μmol·L-1与Pc12细胞预处理12 h,加入LPS 0.15 g·L-1继续培养8 h,采用ccK-8法检测细... 详细信息

目的探讨阿魏酸（FA）对脂多糖（LPS）诱导的Pc12细胞和大鼠海马神经元损伤的保护作用,并探讨其可能的作用机制。方法体外实验：FA 2.5～40μmol·L-1与Pc12细胞预处理12 h,加入LPS 0.15 g·L-1继续培养8 h,采用ccK-8法检测细胞活力;ELISA法检测细胞培养上清中肿瘤坏死因子α（TNF-α）和白细胞介素1β（IL-1β）的释放;激光共聚焦显微镜检测细胞骨架蛋白F肌动蛋白的表达。体内实验：FA25,50和100 mg·kg-1ip给予SD大鼠,每天1次,连续35 d。给药第29天时ip给予LPS 0.2 mg·kg-1,每天1次,连续7d,运用免疫组织化学法观察大鼠海马神经元磷酸二酯酶4B（PDE4B）蛋白表达的变化;Western蛋白质印迹法检测cAMP反应元件结合蛋白（cREB）和磷酸化cREB（p-cREB）表达。结果体内实验：与LPS组细胞比较,FA 10,20和40μmol·L-1预处理组细胞活力明显升高（P〈0.05）,炎性因子TNF-α和IL-1β的释放显著减少（P〈0.05）,细胞骨架蛋白F肌动蛋白的分布和结构明显改善。体内实验：HE染色结果显示,预先给予FA 50和100 mg·kg-1可以减轻LPS诱导的大鼠海马神经元损伤;免疫组织化学实验结果显示,FA 50和100 mg·kg-1能够显著降低LPS诱导的大鼠海马神经元PDE4B蛋白表达水平的升高（P〈0.05）;Western蛋白印迹实验结果表明,FA 50和100 mg·kg-1可逆转LPS对cREB和p-cREB蛋白表达的抑制作用（P〈0.05）。结论 FA对LPS诱导的Pc12细胞和大鼠海马神经元细胞损伤有保护作用,FA抗神经炎症的作用可能与抑制PDE4表达、激活c AMP/cREB信号通路相关。

关键词：阿魏酸磷酸二酯酶4B 脂多糖 c AMP反应元件结合蛋白

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Hepatitis c virus infection and type 1 and type 2 diabetes mellitus

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World Journal of Diabetes 2014年第5期5卷 586-600页

作者： Alessandro Antonelli Silvia Martina Ferrari Dilia Giuggioli Andrea Di Domenicantonio Ilaria Ruffilli Alda corrado Silvia Fabiani Santino Marchi clodoveo Ferri Ele Ferrannini Poupak Fallahi Department of Clinical and Experimental Medicine University of Pisa I-56126 Pisa Italy Department of Medical Surgical Maternal Pediatric and Adult Sciences University of Modena and Reggio Emilia I-41124 Modena Italy Department of Clinical Experimental Medicine University of Pisa I-56126 Pisa Italy Department of Clinical Experimental and Medicine University of Pisa I-56126 Pisa Italy Department of Translational Research and of New Technologies in Medicine and Surgery University of Pisa I-56122 Pisa Italy

Hepatitis c virus(HcV) infection and diabetes mellitus are two major public health problems that cause devastating health and financial burdens worldwide. Diabetes can be classified into two major types: type 1 diabetes mellitus(T1DM) and T2 DM. T2 DM is a common endocrine disorder that encompasses multifactorial mechanisms, and T1 DM is an immunologically mediated disease. Many epidemiological studies have shown an association between T2 DM and chronic hepatitis c(cHc) infection. The processes through which cHc is associated with T2 DM seem to involve direct viral effects, insulin resistance, proinflammatory cytokines, chemokines, and other immunemediated mechanisms. Few data have been reported on the association of cHc and T1 DM and reports on the potential association between T1 DM and acute HcV infection are even rarer. A small number of studies indicate that interferon-α therapy can stimulate pancreatic autoim-munity and in certain cases lead to the development of T1 DM. Diabetes and cHc have important interactions. Diabetic cHc patients have an increased risk of developing cirrhosis and hepatocellular carcinoma compared with nondiabetic cHc subjects. However, clinical trials on HcV-positive patients have reported improvements in glucose metabolism after antiviral treatment. Further studies are needed to improve prevention policies and to foster adequate and cost-effec-tive programmes for the surveillance and treatment of diabetic cHc patients.

关键词： Hepatitis c virus infection Type 1 diabetes mellitus Type 2 diabetes mellitus Epidemiology Pathogenesis Prevention Treatment

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