原核高效制备人乳头瘤病毒16型L1病毒样颗粒

目的通过原核表达系统高效制备人乳头瘤病毒(HPV)16晚期蛋白L1病毒样颗粒(VLPs)。方法构建HPV16L1基因序列优化前后的PET30aHPV16L1重组质粒,转化大肠杆菌Rosetta(DE3);用IPTG诱导目的基因表达,两步层析方法纯化HPV16L1蛋白;电镜下观察纯化产物形成VLPs的情况。结果成功构建大肠杆菌工程菌,高效可溶表达(目的蛋白约占总蛋白的38%)并纯化HPV16L1蛋白,纯度达95%以上,电镜下观察,发现纯化后的目的蛋白为直径50 nm左右,形态与天然病毒颗粒高度相似。结论在大肠杆菌原核系统中高效、简易地制备了HPV16L1VLPs,为诊断试剂和疫苗的研制奠定基础。     

新型溶药器对减少静脉药物配置中不溶性微粒的研究

目的 探讨静脉药物配置的最佳方法,减少静脉输液、静脉注射时不溶性微粒进入体内.方法 百级净化操作台上,采用新型溶药器和注射器分别将氨苄西林钠1、3、6支加入0.9％的氯化钠溶液100ml中,通过微粒分析仪测定两组≥10μm和≥25μm的不溶性微粒,对实验结果进行统计分析.结果 ≥10μm的不溶性微粒溶药器与注射器组间比较差异有显著意义(P＜0.001);≥25μm的微粒加入1支药物后两组比较差异无显著意义(P＞0.05),加入3支药物后两组比较差异有显著意义(P＜0.05);加入6支药物后两组比较差异有显著意义(P＜0.001);各组随着加药数目的增多,溶液中不溶性微粒增多,但溶药器组增加不明显.结论 采用新型溶药器进行药物配置能明显减少不溶性微粒的产生,具有无液体滴漏、操作方便等优点.     

放射性粒子碘-125粒子植入治疗肝癌的围术期护理

目的探讨护理工作在CT导向下放射性粒子植入治疗肝癌中的重要性。方法回顾性分析行碘-125粒子植入治疗的16例原发性肝癌患者临床资料及手术过程。结果所有患者均在精心的护理配合下成功完成CT引导下的碘-125粒子植入术,成功率100%,治疗总有效率为87.5%,无严重并发症发生。结论做好肝癌患者的围术期护理工作,对放射性碘-125粒子植入治疗成功及疗效具有重要意义。     

Effect and mechanism of 125I radioactive particles interposed radiotherapy between organizations on lung cancer

Objective： The aim of this study was to evaluate effect and mechanism of 125I radioactive particles interposed radiotherapy between organizations on lung cancer. Methods： Fourteen cases of patients diagnosed with non-small cell lung cancer （NSCLC）, the use of the B-, CT-guided, according to preoperative imaging and treatment planning system （TPS） program for radioactive particles interposed 125I interstitial radiotherapy. Results： All patients were successfully 125I interstitial radioactive particles interposed radiotherapy. Postoperative local complete tumor remission in 9 cases, partial remission in 5 cases, the efficiency of 100%. No case of serious complications. After 3 to 4 weeks of chemotherapy after 11 cases. 4 cases of lung cancer with bone metastases, pain completely disappeared after treatment. Up to now, five cases have died due to tumor progression, survival time of 12 to 16 months. Nine cases still under follow-up observation and treatment. Conclu- sion： 125I radioactive particles interposed radiotherapy between organizations of lung cancer, simple operation, trauma, fewer complications, conformal high, high local tumor dose, efficacy, and is a supplement of modern radiotherapy techniques for the treatment of lung cancer provides a comprehensive line of the method of effective.     

P物质和PMMA颗粒对成纤维细胞分泌PGE2和IL-6的影响

[目的]在无菌性松动的髋关节假体的界膜发现有P物质免疫活性的感觉神经纤维,本研究探讨P物质和PMMA颗粒对成纤维细胞分泌PGE2和IL-6的影响.[方法]全髋关节置换术后无菌性松动病人翻修术时取松动人工关节周围假滑膜,分离培养成纤维细胞,用不同浓度的P物质、聚甲基丙烯酸甲酯(PMMA)颗粒与成纤维细胞孵育.用酶联免疫分析方法检测细胞培养基中的PGE2和IL-6含量.[结果]P物质或PMMA颗粒能刺激成纤维细胞产生PGE2和IL-6,而且P物质和PMMA颗粒共同刺激的成纤维细胞产生PGE2和IL-6明显增加.[结论]P物质和PMMA颗粒在增强成纤维细胞PGE2和IL-6的分泌方面有协同作用.PGE2和IL-6在骨溶解和假体松动的发展过程中起重要作用.因此,PMMA颗粒和P物质都能在界膜的炎症、骨溶解和关节假体无菌性松动中起作用.     

清胰颗粒对重症急性胰腺炎大鼠肠黏膜上皮紧密连接的保护作用

目的：探讨清胰颗粒对重症急性胰腺炎（SAP）大鼠肠黏膜上皮紧密连接的保护作用。方法：24只健康雄性Wistar大鼠随机分为假手术组、SAP组和清胰颗粒组。采用3.5%牛磺胆酸钠胆胰管逆行注射制备SAP大鼠模型,清胰颗粒组于造模前2 h、造模后6 h和18 h分别给予清胰颗粒灌胃,SAP组以同样的方法给予安慰剂。各组分别于造模后24 h取末端回肠,HE染色,光镜下观察病理组织学改变,采用Chiu’小肠组织损伤评价标准对肠黏膜损伤进行评价,应用Western blot和实时荧光定量PCR法检测回肠黏膜上皮紧密连接蛋白（Occludin）表达。结果：与假手术组相比,SAP组大鼠Chiu’小肠组织损伤评分（4.6±0.6） vs （0.0±0.0）明显升高,与SAP组相比,清胰颗粒组Chiu’小肠组织损伤评分（2.0±0.4）vs（4.6±0.6）明显降低;Western blot和实时荧光定量PCR结果显示,与假手术组相比SAP组大鼠Occludin蛋白表达（1.2±0.4）vs（3.2±0.4）和mRNA表达（1.3±0.3）vs（2.9±0.5）明显降低（P＜0.05）;与SAP组相比,清胰颗粒组 Occludin蛋白表达（4.1±0.4）vs（1.2±0.4）和mRNA表达（4.2±0.3）vs（1.3±0.3）明显升高（P＜0.05）。结论：清胰颗粒能减轻SAP大鼠肠黏膜损伤程度,这一作用可能与增强肠黏膜紧密连接蛋白Occludin表达有关。     

登革2型病毒包膜蛋白在毕赤酵母中的表达和病毒样颗粒的组装

目的在组成型的毕赤酵母系统中表达登革2型病毒包膜蛋白E和前膜蛋白prM,并研究病毒样颗粒在酵母细胞中的组装特性。方法以登革2型病毒ZS01/01株mRNA为模板,扩增编码prM/E蛋白的基因。将目的基因克隆至表达载体pGAPZaA的多克隆位点,并转化毕赤酵母GS115。经Zeocin抗生素筛选和PCR鉴定后获得酵母菌重组克隆,并进行表达和鉴定。结果成功克隆登革2型病毒prM/E基因,SDS-PAGE和Western blot检测表明包膜蛋白E在组成型的毕赤酵母系统中得到高水平表达;利用透射电镜在重组酵母胞浆内检测到30-50 nm的登革2型的病毒样颗粒,并发现类似登革病毒感染的双层膜结构;免疫电镜结果证实重组病毒样颗粒可以与抗登革2型病毒血清反应,具有免疫反应性。结论成功的建立了组成型表达登革2型病毒样颗粒的毕赤酵母系统,为登革病毒样颗粒疫苗的制备奠定了基础。     

吸尘罩的优选及其在玉石雕刻防尘中的应用

目的对适用吸尘罩进行优选,并在实际通风除尘系统中应用。方法用示踪气体法对吸尘罩进行优选,比较不同吸尘罩捕集效率和风量的关系;对吸尘罩前风速分布进行测定,得出罩前风速分布模型。并对吸尘罩进行空气动力性能分析,测定雕刻排气中粉尘的粒径分级组成和除尘器的粉尘粒径分级除尘效率,评估除尘器的总除尘效率。结果优选出的雕刻吸尘罩,置于切割轮正前方0.11 m处,其控制风速为0.5 m/s,能有效地控制雕刻切割轮的粉尘不向周围扩散,工作地点空气粉尘浓度由无罩时的30-297 mg/m3降低至1.1-1.7 mg/m3,吸尘罩风量为255 m3/h,为其他类型吸尘罩的50%-77%。优选出的冲击水浴除尘器,其除尘效率为97%-98%,阻力为1.5 kPa;滤袋除尘器的效率为98%-99%,阻力为2.5 kPa;除尘后尾气的粉尘浓度为20-30 mg/m3。结论提出了优选吸尘罩的设计方法及实用的除尘器,可供实际应用。     

Characterization of process air emissions in automotive production plants

During manufacturing, particles produced from industrial processes become airborne. These airborne emissions represent a challenge from an industrial hygiene and environmental standpoint. A study was undertaken to characterize the particles associated with a variety of manufacturing processes found in the auto industry. Air particulates were collected in five automotive plants covering ten manufacturing processes in the areas of casting, machining, heat treatment and assembly. Collection procedures provided information on air concentration, size distribution, and chemical composition of the airborne particulate matter for each process and insight into the physical and chemical processes that created those particles.     

Inhibition of the binding of HCV serum particles to human hepatocytes by E1E2‐specific D32.10 monoclonal antibody

The aim of this study was to determine the inhibition of binding activity of the monoclonal antibody (mAb) D32.10 which recognizes a highly conserved discontinuous antigenic determinant (E1:297–306, E2:480–494, and E2:613–621) expressed on the surface of serum‐derived HCV particles (HCVsp) of genotypes 1a, 1b, 2a, and 3a. To this end, an in vitro direct cell‐binding assay based on the attachment of radiolabeled HCVsp was developed, and Scatchard plots were used to analyze ligand–receptor binding data. HCV adsorption was also assessed by quantitating cell‐associated viral RNA by a real‐time RT‐PCR method. Saturable concentration‐dependent specific binding of HCVsp to Huh‐7 or HepaRG cells was demonstrated. The Scatchard transformed data showed two‐site interaction for Huh‐7 and proliferative HepaRG cells: the high‐affinity binding sites (K_d1 = 0.1–0.5 µg/ml) and the low‐affinity binding sites (K_d1 = 5–10 µg/ml), and one‐site high‐affinity binding model between E1E2/D32.10‐positive HCVsp and hepatocyte‐like differentiated HepaRG cells. The E1E2‐specific mAb D32.10 inhibited efficiently (>60%) and selectively the binding with an IC₅₀ ≤0.5 µg/ml in all the experimental approaches using serum HCV of genotype either 3 or 1b. This supports the involvement of the E1E2/D32.10 discontinuous antigenic determinant in the interactions between human hepatocytes and HCVsp, and suggests that D32.10‐like antibodies present in sera from patients infected with HCV could play a protective role. J. Med. Virol. 81:1726–1733, 2009. © 2009 Wiley‐Liss, Inc.