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

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

总责任护士主导的医护一体化护理模式在创伤骨科的应用

目的探讨总责任护士主导的医护一体化临床护理模式在创伤骨科的应用效果。方法选择2016年7月至2017年6月在南京大学医学院附属鼓楼医院创伤骨科行手术治疗的骨折患者200例,按照入院先后顺序分为对照组90例(实施创伤骨科常规护理)和观察组110例(实施医护一体化工作模式,包括成立医护合作小组、实施总责任护士主导的医护一体化查房、个体化诊疗方案的共同决策、疼痛管理、早期出院计划与延续护理)。比较2组患者的术后护理效果和医护合作满意度。对数据行t检验和χ~2检验。结果总责任护士主导的医护一体化模式下,观察组患者术后72 h内数字评分法(NRS)评分为(4.09±1.64)分,疼痛干预有效率为77.57%,对照组术后72 h内NRS评分为(5.15±1.69)分,疼痛干预有效率为57.95%,2组比较差异均有统计学意义(P值均小于0.05);观察组下肢深静脉血栓、泌尿系统感染、便秘的发生例数分别为2、 1、3,对照组下肢深静脉血栓、泌尿系统感染、便秘的发生例数分别为9、7、11,两组比较差异均有统计学意义(χ~2=4.486、4.296、6.812,P=0.027、0.036、0.009)。观察组术后康复开始时间为(21.48±6.41)h,术后住院时间为(11.08±2.19)d;对照组术后康复开始时间为(36.48±10.75)h,术后住院时间为(13.95±2.93)d,2组比较差异均有统计学意义(t=11.670、-7.616,P值均小于0.05)。2组医护合作满意度结果显示,观察组患者信息的交流得分为(3.78±0.85)分,共同参与治疗或护理决策得分为(3.86±0.87)分,医师和护士的关系得分为(4.00±0.84)分,总分为(3.87±0.67)分;对照组患者信息的交流得分为(3.32±1.00)分,共同参与治疗或护理决策得分为(3.23±0.93)分,医师和护士的关系得分为(3.45±0.96)分,总分为(3.33±0.84)分,2组比较差异均有统计学意义(P值均小于0.05)。结论总责任护士主导的医护一体化护理模式的应用,以亚专科为核心,重新整合医护资源,共同参与制订患者的诊疗护理方案,共同聚焦患者安全与质量,加强了医护间的配合,提高了医疗护理服务质量,提升医护合作满意度。国家自然科学基金面上资助项目(81372051)北京市科技计划“首都特色”专项(Z151100004015199

X连锁淋巴细胞异常增生症死亡1例

XLP存在复杂多样的临床表现,其中以爆发性传染性单核细胞增多症型临床经过尤为凶险,诊疗过程中多无法及时诊断,且对症治疗方案多无明显效果,患者可迅; 速死亡,极易引起医疗损害而需要法医学鉴定。笔者认为遇到怀疑XLP进行鉴定时需把握下列几个要点:(1)该病发病机理为病毒感染引起淋巴细胞过度增殖,; 浸润多个器官,需做CD20、CD3、TdT、MPO等抗体的免疫组织化学染色,以鉴定增殖淋巴细胞的类型,排除其他血液系统疾病;(2)该疾病属性连锁; 隐性遗传性疾病,应尽量询问、调査被鉴定人相关亲属的资料,以确定其是否与遗传相关,须与散发性传染性单核细胞增多症鉴别;(3)由于基层法医鉴定机构多; 不具备相关分子生物学检査手段,无法从基因突变上作出明确诊断,还需依据尸体检验结果并结合具体临床表现综合考虑

气管旁淋巴结结核侵及气管和头臂干动脉致死1例

1案例1.1简要案情张某,女,35岁,某年6月17日,以"反复咳嗽、咳痰、气喘1月余"为主诉入院。入院诊断:前纵隔占位性质待查,气管狭窄。入院后完善检查,于6月20日行支气管镜检查,见气管中段严重狭窄,表面见扁平新生物并溃烂,取新生物送检并置入Y型硅酮支架。6月26日17:00张某无明显诱因下突发大咯血,咯血量约1 000 mL,之后迅速出现意识丧失,给予抢

11例脂肪栓塞法医学鉴定分析

目的探讨脂肪栓塞法医学鉴定的思路及注意要点。方法回顾性分析某司法鉴定中心2007年1—12月共计11例脂肪栓塞死亡的案例,观察HE切片及苏丹IV染色切片。结果 11例肺组织内均见大量脂滴,2例脑和其他脏器小血管内见脂滴,肝脂肪变性6例。结论结合案例分析脂肪栓塞栓子来源,临床表现和死亡机制,总结出脂肪栓塞案件鉴定的一些经验和工作思路

Retrofitting of deep concrete coupling beams by laterally restrained side plates

published_or_final_versionCivil EngineeringDoctoralDoctor of Philosoph

Study of Dynamic Characteristics of Cylinder Profile of Rotary Vane Compressor for Automotive Air Conditioner

Cylinder Profile is one of the key factors influencing the performance of rotary vane compressor used for automobile air conditioning system. A new type of cylinder profile is put forward in the paper, which is composed of three parts: circle arc near the minor axis of cylinder profile, main curve and transit curve between them. Volume ratio of the compressor, motion of vane, forces and moments acting on the rotor are studied with some typical cylinder profiles. It is shown that the profile composed of trigonometric function has better velocity and acceleration characteristics than other profiles mentioned in the paper. Stress under impact and noise pollution of the compressor could be reduced effectively by optimization design of the transit curve. Cylinder profile has little influence on shaft power of rotary vane compressor

Study of Dynamic Characteristics of Cylinder Profile of Rotary Vane Compressor for Automotive Air Conditioner

Cylinder Profile is one of the key factors influencing the performance of rotary vane compressor used for automobile air conditioning system. A new type of cylinder profile is put forward in the paper, which is composed of three parts: circle arc near the minor axis of cylinder profile, main curve and transit curve between them. Volume ratio of the compressor, motion of vane, forces and moments acting on the rotor are studied with some typical cylinder profiles. It is shown that the profile composed of trigonometric function has better velocity and acceleration characteristics than other profiles mentioned in the paper. Stress under impact and noise pollution of the compressor could be reduced effectively by optimization design of the transit curve. Cylinder profile has little influence on shaft power of rotary vane compressor