Inhibitory effect of anti-human NRP-1 monoclonal antibody on hepatocellular carcinoma cell line HepG2 and its mechanism in vitro

目的研究抗人神经纤毛蛋白-1(Neuropilin1,NRP-1)单克隆抗体对肝癌Hep G2细胞的生长抑制作用及其机制。方法小鼠腹水法制备抗NRP-1单克隆抗体(NRP-1 m Ab)并用r Protein A亲和柱纯化抗体,间接ELISA检测抗体的滴度水平。Western blot检测NRP-1 m Ab对Hep G2细胞的特异性,细胞免疫荧光和流式细胞术检测NRP-1蛋白在肝癌细胞株Hep G2上的表达,MTT法检测NRP-1 m Ab对Hep G2的生长抑制作用,Western blot检测ERK1/2、P-ERK1/2、Akt、P-Akt蛋白的表达水平。结果 SDS-PAGE和间接ELISA检测纯化的NRP-1 m Ab纯度为95%以上,效价为1×10~(-6);Western blot检测结果显示NRP-1 m Ab可与Hep G2细胞膜上的NRP-1蛋白特异性结合。细胞免疫荧光染色结果显示NRP-1定位于Hep G2细胞膜,流式细胞术结果显示NRP-1蛋白在Hep G2细胞上表达水平较高;MTT法检测结果显示NRP-1 m Ab对Hep G2细胞有生长抑制作用。Western blot检测到在不同浓度NRP-1 m Ab作用下,Hep G2细胞裂解液P-ERK1/2、P-Akt蛋白的条带信号逐渐减弱。结论纯化的NRP-1m Ab能抑制Hep G2细胞的生长,其抑制作用是通过EGF和HGF信号通路实现的。The aim of the experimental is to investigate the inhibitory effect of anti-human nerve cilia protein1(Neuropilin-1,NRP-1) monoclonal antibody(NRP-1 mAb) on hepatocellular carcinoma cell line HepG2 and its mechanism in vitro.Anti-human NRP-1 monoclonal antibody(NRP-1 mAb) was prepared from mouse ascites and purified by rProteinA affinity column assay.The titer of antibody was determined using indirect ELISA assay;the characteristic of NRP-1 mAb binding to NRP-1 was determined using Western blotting;the expression of NRP-1protein in hepatocellular carcinoma cell line HepG2 was determined using immunofluorescence assay and flow cytometry assay.Growth inhibition of HepG2 cells treated with different concentrations of NRP-1 mAb was determined using MTT assay,while Western blotting was used to detect the expression levels of ERK1/2,P-ERK1/2,Akt and P-Akt proteins.The results of SDS-PAGE and indirect ELISA showed that the purity of purified NRP-1mAb was more than 95%and the titer was 1×10~(-6).Western blotting analysis suggested that NRP-1 mAb could bind specifically to NRP-1 on HepG2 cell;immunofluorescence staining showed that NRP-1 was located in the membrane of HepG2 cells.Flow cytometry analysis showed that the expression level of NRP-1 on HepG2 cell was relatively high.Western blotting analysis suggested that P-ERK1/2 and P-Akt expression levels were down-regulated after having incubated HepG2 cells with different concentrations of NRP-1 mAb.In conclusion,NRP-1 mAb could inhibit the growth of HepG2 cells(P<0.05),and its inhibitory effect is achieved by reducing the P-ERK1/2 and P-Akt expression.南京军区医学科技创新项目(12MA061,15MS104

The Determination of Polycyclic Aromatic Hydrocarbons in Sediments with ASE/GC-MS

建立了用加速溶剂萃取技术提取含硫沉积物中痕量多环芳烃的可靠方法,优化了不同萃取剂,不同提取程序等萃取条件,研究了沉积物中的硫化物对色谱分离的影响。15种多环芳烃的检出限为0.09~1.34ng/g,相对标准偏差(RSD,n=6)为12.5%~23.9%,15种多环芳烃的低浓度加标回收在74%~138%之间。高浓度加标回收率除吲哚芘为62%外,其余都在80%~122%之间。基本上达到了痕量分析的要求。用该方法分析山东半岛近岸海域沉积物中多环芳烃的含量,测得15种PAHs的总量为273~579 ng/g。与其它地区沉积物中多环芳烃含量相比,属轻/中度污染。An accelerated solvent extraction method for the extraction of polycyclic aromatic hydrocarbons(PAHs) in sediments was developed.The extraction conditions such as extraction solvent and program were optimized.The effects of sulfide on chromatographic separation were studied.The detection limits for the fifteen PAHs were 0.09-1.34 ng/g,and the relative standard deviations(RSDs) ranged from 12.5% to 23.9%.The recoveries of high level concentration ranged from 62% to 122% and those of low level concentration ranged from 74%-138%.The contents of PAHs in sediment samples collected in the coastal regions off the Shandong peninsula,China were analyzed by this method.The PAHs in these samples ranged from 273 ng/g to 579 ng/g.Overall,the PAHs levels were comparable to those reported in the literature for sediment samples collected from similar environments around the world.国家高技术研究发展计划(863)基金项目(2003AA635180);; 青岛科技将才计划基金项目(04-3-J.J-11

固相微萃取-气相色谱-质谱联用测定海水和沉积物间隙水中的痕量多环芳烃

建立了固相微萃取(SPME)与气相色谱-质谱(GC-MS)联用同时测定海水中16种多环芳烃的分析方法,研究了萃取时间、盐度条件的影响．同时用SPME的方法研究了海水中的溶解有机物(DOM)对多环芳烃萃取的影响．计算出不同DOM浓度下多环芳烃KDOM与KOW的关系: CDOM=5 mg／L时,logKDOM=0．7944KOW+0．773(R2=0．91)．CDOM=10 mg／L时,logKDOM= 0．7905KOW+0．668(R2=0．97);CDOM=30 mg／L时,logKDOM=0．714KOW+1．0407(R2=0．91)．该法对16种多环芳烃的检出限为0．1-3．5 ng／L,相对标准偏差(RSD,n=5)为4％-23％．用该法分析海洋环境中的痕量多环芳烃,16种多环芳烃的平均回收率为88.2±20．4％,方法快速、灵敏、简单,适用于快速分析海水和沉积物间隙水样中的痕量多环芳烃．国家高技术研究发展计划(863)(批准号:2003AA635180);; 青岛科技将才计划(批准号:04-3-JJ-1)资助项

重组人血管内皮生长抑制因子对人肺腺癌裸鼠移植瘤的放射效应影响

目的:探讨重组人血管内皮抑制因子(recombinant human vascular endothelial growth inhiloitor-192,rhVEGI-192)对人肺腺癌裸鼠移植瘤的放射增敏作用。方法:采用原核表达rhVEGI-192,获得目的蛋白。通过肿瘤倍增时间,计算药物的增敏系数。通过建立人肺腺癌裸鼠移植瘤模型,荷瘤裸鼠随机分为4组:对照组、10Gy、rhVEGI-192、rhVEGI-192+10Gy。采用6MV-X线进行照射,照射剂量为10Gy。获得移植瘤标本,利用免疫印迹法检测移植瘤中VEGF(vascular endothelial growth factor)的表达变化。结果:SDS电泳结果显示,目的蛋白位于22k D左右。10Gy照射时,重组人血管内皮抑制因子的EF(enhancement factor)值为1.5。和空白对照组相比,rhVEGI-192组和10Gy组移植瘤的生长受到抑制(P<0.001),rhVEGI-192+10Gy组移植瘤生长显著抑制(P<0.001),rhVEGI-192+10Gy组移植瘤较10Gy组有明显生长抑制。和空白组相比,rhVEGI-192组VEGF表达减少,而10Gy组VEGF表达变化不明显,rhVEGI-192+10Gy组VEGF表达明显减少。rhVEGI-192+10Gy和rhVEGI-192组相比,VEGF表达减少。结论:rhVEGI-192联合照射能够减少VEGF的表达。这可能是rhVEGI-192的增敏机制之一。中国人民解放军南京军区医学科技创新项目(编号:No.12MA061

ONE NEW GENUS,TWO NEW SPECIES AND ONE NEW RECORD of CORYMORPHIDAE FROM THE SOUTH CHINA SEA(ANTHOMEDUSAE,CAPITATA)

记述了采自南海的棒状水母科COryMOrPHIdAE AllMAn,18721新属,肋突水母属COSTA HuAng,Xu ET lIn,gEn.nOV.,2新种,即南海肋突水母COSTA nAnHAIEnSIS HuAng,Xu ET lIn,SP.nOV.,间腺梅尔水母MAyErI InTErgOnA HuAng,Xu ET guO,SP.nOV.和1新纪录种,幼芽真囊水母EuPHySOrA gEMMIfErA bOuIllOn,1978。详细描述其形态特征,并绘图与本属其它种进行比较,模式标本保存于国家海洋局第三海洋研究所。Samples of Corymorphidae were collected from the South China Sea(4°-18° 30' N,109° 119°E),during Aug.2011.Based on the taxonomic system of Bouillon et al.(2006),at present time 12 valid genera of Corymorphidae are known,Euphysora M aas,1905 and Vannuccia Brinckmann-Voss,1967 are valid genera.In view of the used of the genus name of Vannuccia M arcus,1948,we wish to make the proposal of which the genus name of Vannuccia BrinckmannVoss,1967 is change to that of Mayeri Xu,Huang et Guo,gen.nov.,remembering the first author for the species of the genus.One new genus,Costa Huang,Xu et Lin,gen.nov.,two new species,i.e.Costa nanhaiensis Huang,Xu et Lin,sp.nov.,Mayeri intergona Huang,Xu et Guo,sp.nov.and one new record,Euphysora gemmifera Bouillon,1978 are described.All type specimens are deposited in the T hird Institute of Oceanography,SOA.T he specific diagnoses of one new genus and two new species are as follows.Costa Huang,Xu et Lin,gen.nov.Umberlla dome-shaped,exumbrella with 4 longitudinal ridges and ribs;manubrium large,the length no extending umbrella margin;mouth simple,circular;one well-developed solid tentacle terminating in large cnidocyst knob,3 marginal bulbs without tentacles;4 radial canals,1 circular canal;with mesenteries.T ype species:Costa nanhaiensis Huang,Xu et Lin,sp.nov.Etymology.T he genus name from Latin Costa,means rib and ridge,the new genus exumbrella with 4 longitudinal ridges and ribs.Costa nanhaiensis Huang,Xu et Lin,sp.nov.(Figs 1-2) Umbrella dome-shaped,exumbrella with 4 longitudinal ridges and ribs;manubrium large,extending no beyond umbrella margin,vacuolated endodermal cells in the upper part,gonads in the lower part;one well-developed solid tentacle terminating in two large cnidocyst knobs;3 marginal bulbs without tentacles;4 radial canal,1 circular canal;with mesenteries.Etymology.Form Latin,Nanhaiensis,means Nanhai,the species name refers to the first collected from Nanhai(the South China Sea).Mayeri intergona Huang,Xu et Guo,sp.nov.(Figs 3-4) Umbrella dome-shaped,without apical process,bell margin slightly asymmetrical;manubrium short,length about 1 /3 of the umbrella cavity,with a apical chamber;gonads global shaped,in interradial position of manubrium;one swollen marginal tentacle,hollow for half its length and ending in long,large,clubshaped swelling armed with cnidocysts.我国近海海洋综合调查与评价(908专项);“中国海洋生物种类名录和图谱”(908-ZC-Ⅱ-02);“908专项海洋生物样品库”和“全球变化与海气相互作用专项”资助---

TWO NEW SPECIES of THE GENUS NUBIELLA FROM THE SOUTH CHINA SEA(FILIFERA,BOUGAINVILLIDAE)

记述了采自南海花水母亚纲AnTHOMEduSAE HAECkEl,1879、丝螅水母目fIlIfErA küHn,1913、高手水母科bOugAIn VIllIdAE lüTkEn,1850、单肢水母属nubIEllA bOuIllOn,19802新种,即球形单肢水母n. glObOSA lIn,Xu ET HuAng,SP.nOV.和球腺单肢水母n. glObOgOnA WAng,guO ET XuE,SP.nOV.,对新种特征进行了绘图和描述。模式标本保存于国家海洋局第三海洋研究所。Samples of Nubiella were collected from the South China Sea(4°-18° 30' N,109°-119° E) during Aug.2011.T wo new species of Nubiella i.e.N.globosa Lin,Xu et Huang,sp.nov.and N.globogona W ang,Guo et Xue,sp.nov.are described.All type specimens are deposited in the T hird Institute of Oceanography,SOA.T he specific diagnoses of two new species are as follow.Nubiella globosa Lin,Xu et Huang,sp.nov.(Figs 1-2) Umbrella global-shaped,without apical projection,exumbrella without scattered cnidocysts;without gastric peduncle and oral tube;gonads on interradial region of manubrium;with 4 unbranched oral tentacles;4 radial canals thin;4 tentacular bulbs global-shaped,without dense masses;4 marginal tentacles rather short with ring cnidocysts.Nubiella globogona Wang,Guo et Xue,sp.nov.(Figs 3-4) Umbrella bell-shaped,without apical projection,exumbrella with scattered cnidocysts;manubrium cylindrical-shaped,with apical chamber;mouth simple,circular;with 12 unbranched oral tentacles;gonads globular-shaped on media interradial region of manubrium;4 marginal tentacles,tentacular bulbs with black pigments.我国近海海洋综合调查与评价(908专项);“中国海洋生物种类名录和图谱”(908-ZC-Ⅱ-02);“908专项海洋生物样品库”和“全球变化与海气相互作用专项”资助---

Application of oil spill identification by GCMS——A case study

通过对中国近海海域一个具体海面溢油事例的研究,提出了通过气相色谱及气质联用技术进行海面溢油鉴别的方法。采用GC-FID对油样中的正构烷烃的主要组分进行定性分析,利用GC-MS对油中难降解的生物标记化合物:霍烷及其他五环三萜类化合物、规则甾烷及重排甾烷类化合物、三芳甾类化合物、多环芳烃进行分析,计算相应的比值并采用"t-分布"方法进行溢油样品和可疑溢油源样的相关分析。结果如下:(1)从GC-FID色谱图可以看出,YY2679与KY0631、KY0632的色谱图极其相似,与其余四个可疑油源的色谱图差异较大。六个可疑油源的R值在0.04~1.95之间,只有KY0631和KY0632的R值小于0.1,KY0633~KY0636与YY2679的差异不是由风化引起的,可以排除,由此可以推断出YY2679溢油样品可能来源于KY0631或KY0632两个可疑溢油源。(2)采用21个生物标志物指标进行相关性判别,KY0631可疑溢油源在比值点的区间范围为95%和98%时,有多个点x或y误差棒均未跨过直线y=x。KY0632可疑溢油源在比值点的区间范围为95%时,各点的x或y误差棒均跨过直线y=x,由此推断YY2679溢油样品和可疑油源KY0632是同一油源。On the basis of a case study,a method to identify the spilled oil at sea was introduced.N-alkane distribution patterns of unknown oil were recognized by gas chromatography-mass spectrometry determined the distribution of a suit of petroleum biomarkers hopanes,triterpanes,regular steranes diasteranes,triaromatic steroids and polycyclic aromatic hydrocarbons.The calculated diagnostic ratios for selected biomarker in the source of the spilled oil were done.The correlation studies compared the spill sample and suspected source were made.The results showed three samples nearly identical GC chromatographic profiles,n-alkane distribution patterns,as well as nearly identical diagnostic ratios.This implies that they were most likely from the same source.In 95%confidence limit,all diagnostic ratios of sample KY0632 is exactly the same as for YY2679,so conclusion that the sample of KY2679 and YY0632 are come from the same source

ST段抬高型急性心肌梗死院前溶栓治疗中国专家共识

急性心肌梗死仍然严重威胁我国人民健康,在我国广大城乡地区,形势更为严峻[1,2]。及时救治急性心肌梗死患者,降低死亡率和保护心脏功能刻不容缓。鉴于我国的实际情况,院前溶栓治疗在大城市以外的城乡地区具有重要意义。为此,中国医师协会胸痛专业委员会及中国医学救援协会心血管急救分会专门组织有关专家制订了本共识,旨在帮助院前医疗急救人员对急性心肌梗死患者选择最佳

长汀县赢坪史前遗址发掘简报

2012年福建博物院对赢坪两处遗址进行了抢救性考古发掘,获得了一批包括石器、陶器、铜器、石范等内涵丰富的文化遗物,这批遗物可进一步了解闽西地区的史前文化发展序列及文化面貌,并且对于多地区文化间的交流研究也有一定的作用