两亲性聚乙烯亚胺修饰量子点构建功能性光学探针用于干细胞治疗的示踪研究

目的：制备新型分子影像探针标记间充质干细胞,以实现细胞荧光成像。方法应用1‐碘十二烷修饰聚乙烯亚胺,构建两亲性大分子。利用大分子自组装性能,包裹疏水性核壳结构CdSe/CdS量子点构建纳米探针。结果该探针在630 nm处呈现出较强的荧光信号,能够促使间充质干细胞高效内吞。体内成像结果显示,与空白对照细胞相比,被标记细胞呈现出显著的光学信号,空白组信号值为1.47 e8[p/s/cm2/sr]/[μW/cm2],实验组信号值为2.78 e8[p/s/cm2/sr]/[μW/cm2]。结论该纳米探针具有良好光学成像性能,可实现细胞标记。     

Dot enzyme‐linked immunosorbent assay strip as a screening tool for detection of autoantibody to interferon gamma in sera of suspected cases of adult‐onset immunodeficiency

Background

Being able to detect the presence of autoantibodies to interferon (IFN)‐γ in serum is essential for evaluating patients with suspected adult‐onset immunodeficiency (AOID) with unusual intracellular infections. Most reported patients with AOID have been Asian, although the exact prevalence of this illness is unknown. To date, no standard assay exists to detect autoantibodies to IFN‐γ. An easy‐to‐use, low‐cost assay that can be performed in any laboratory would be a valuable tool for clinical management of AOID, as well as better reveal its prevalence.

Methods

Our experimental study exploited a dot enzyme‐linked immunosorbent assay (Dot‐ELISA) strip to detect autoantibodies to IFN‐γ. Sera from 66 HIV‐negative patients having autoantibodies to IFN‐γ as determined by indirect ELISA were tested.

Results

Dot enzyme‐linked immunosorbent assay was sensitive (100%) and specific (94.5%), with a positive predictive value of 97.6% and a negative predictive value of 100%.

Conclusion

This simple method provides prompt qualitative results that can be read visually and used in facilities with limited testing capabilities.

     

基于计算机仿真技术的中药水提液中药效物质与共性高分子物质“溶液结构”及相互作用初探——以黄连解毒汤为例

目的：分析淀粉、果胶等共性高分子物质与小分子药效物质的共存状态及相互作用。方法：选择黄连解毒汤模拟液为研究对象,使用计算机仿真技术,对分析物质进行建模、体系能量最小化、计算总体系能量、计算共性高分子单独存在时的能量、计算小分子药效物质单独存在时的能量,分析淀粉、果胶等共性高分子物质与小檗碱、巴马丁、黄芩苷、栀子苷的相互作用关系。结果：淀粉、果胶与小檗碱、巴马汀为相互吸引关系,与黄芩苷为弱相互吸引关系,而与栀子苷则为相互排斥作用;淀粉与小檗碱、巴马丁、黄芩苷、栀子苷的平均相互作用能依次为-32.159,-28.630,-5.974,234.295 kcal·mol^-1,果胶与小檗碱、巴马丁、黄芩苷、栀子苷的相互作用能平均值分别为-19.717,-19.804,-1.067,253.258 kcal·mol^-1;淀粉、果胶和小分子药效物质的分离容易顺序依次为栀子苷 >黄芩苷 >巴马汀 >小檗碱。结论：共性高分子和小分子药效物质间的相互作用由范德华力占主导地位,静电作用较小（3%~15%）,相互作用能的大小和正负值与药效物质的空间位置有关。     

Identification of Old World Leishmania species by PCR–RFLP of the 7 spliced leader RNA gene and reverse dot blot assay

The aim of the study was to assess the 7SL RNA PCR followed by restriction fragment length polymorphism (RFLP) and reverse dot blot (RDB) assays for use in identification of Old World Leishmania species. Species‐specific RFLP patterns were obtained for Leishmania major, Leishmania tropica and the Leishmania donovani complex when the 7SL RNA PCR product was digested with the restriction enzyme BsuRI, an isoschizomer of HaeIII. For the RDB assay, biotin‐labelled 7SL RNA amplicons were hybridized to Leishmania genus‐specific and species‐specific oligonucleotide probes immobilized onto a membrane. The Old World Leishmania species could be distinguished by using five probes: one that was a genus‐specific probe and hybridized to all Leishmania species (Lc), two that were specific for L. major (Lm1 and Lm2), one that was specific for L. tropica (Lt) and one that detected both L. major and L. tropica (Lmt). The PCR–RDB was 10 times more sensitive than 7SL PCR and can detect <1 parasite. In addition, the identification of species was easier and more reliable than with 7SL PCR–RFLP. 7SL PCR–RFLP detected parasites in 50 of 57 clinical samples, whereas PCR–RDB detected 53 and 55 were detected by amplification of kinetoplast (k) DNA. The 7SL RNA PCR has proven useful for direct diagnosis of Old World leishmaniasis, especially when combined with the RBD assay for species identification.     

间歇性外斜视手术量与术前立体视状况相关性研究

目的探讨间歇性外斜视术前立体视状况与手术量的关系。方法回顾分析47例常规手术量矫正的间歇性外斜视患者的术前远近立体视觉及术后眼位,与16例术前有远立体视并采用调整手术量患者手术后眼位对比研究。结果回顾性资料中,术前无立体视觉和只有近立体视觉的患者,其术后正位率,二者差异无统计学意义(P>0.05)。而术前有远立体视者,其术后正位率与术前无立体视及只有近立体视者差异均有统计学意义(P<0.01)。研究资料中,术前有远立体视觉的26例患者均在常规手术量的基础上适当减少手术量,术后眼正位率,与术前无立体视及仅有近立体视而采用常规手术量的患者相比较差异均无统计学意义(P>0.05)。结论术前有远立体视的间歇性外斜视,适当减小手术量,不影响手术成功率。     

槲皮素对人食管癌Eca-109细胞的分化诱导作用

目的探讨槲皮素对人食管癌Eca-109细胞的分化诱导作用。方法将培养的Eca-109细胞分为两组:①加槲皮素组(Q组);②不加药物对照组(C组)。同时培养48h,收集各组细胞制备成细胞硝酸纤维素膜(NCM)标本及提取RNA制备RNA硝酸纤维素膜标本。分别对细胞硝酸纤维素膜(NCM)标本进行PCNA,VEGF的免疫斑点印迹阵列;对RNA硝酸纤维素膜标本EGFR,c-myc及wtp53的RNA斑点印迹阵列。印迹阵列皆用岛津薄层色谱扫描仪(TLC)进行波长523nm的扫描数值比较。结果①免疫斑点印迹显示:Q组的PCNA-IR,VEGF-IR的TLC扫描数值分别比C组低5.5倍,3.5倍;②RNA斑点印迹阵列显示,与C组相比:Q组的EGFR,c-myc的RNA斑点TLC扫描数值分别下调了7倍,2.2倍。而wtp53的RNA斑点TLC扫描数值则上调了2.2倍。结论槲皮素可下调作为肿瘤细胞恶性程度生物学指标的PCNA,EGFR,VEGF,c-myc等的表达,上调抑癌基因wtp53的表达,表明对人食管癌Eca-109细胞具有一定的诱导分化效应。     

Lipid peroxidation induced by indomethacin with horseradish peroxidase and hydrogen peroxide: involvement of indomethacin radicals

Some of the side-effects of using indomethacin (IM) involve damage to the gastric mucosa and liver mitochondria. On the other hand, neutrophils infiltrate inflammatory sites to damage the tissues through the generation of reactive oxygen species by myeloperoxidase. The stomach and intestine have large amounts of peroxidase. These findings suggest that peroxidases are involved in tissue damage induced by IM. To clarify the basis for the tissue damage induced by IM in the presence of horseradish peroxidase (HRP) and H2O2 (HRP-H2O2), lipid peroxidation was investigated. When IM was incubated with liver microsomes in the presence of HRP-H2O2 and ADP-Fe3+, lipid peroxidation was time-dependent. Catalase and desferrioxamine almost completely inhibited lipid peroxidation, indicating that H2O2 and iron are necessary for lipid peroxidation. Of interest, superoxide dismutase strongly inhibited lipid peroxidation, and it also inhibited the formation of bathophenanthroline-Fe2+, indicating that reduction of the ferric ion was due to superoxide (O2-). ESR signals of IM radicals were detected during the interaction of IM with HRP-H2O2. However, the IM radical by itself did not reduce the ferric ion. These results suggest that O2- may be generated during the interaction of IM radicals with H2O2. Ferryl species, which are formed during the reduction of iron by O2-, probably are involved in lipid peroxidation.     

在屈光习惯矫正与全矫状态下用Worth四点法与卡洞法测量优势眼的结果比较

目的研究Worth四点法测量优势眼与卡洞法测量优势眼的相关性,并分别研究这两种方法在习惯矫正情况下与屈光全矫状况下测量优势眼眼别的差异。方法429例受试者,其中部分受试者采用两种方法(卡洞法与Worth四点法)测量优势眼,部分受试者接受了在两种条件下(习惯矫正方式与屈光全矫下)测量优势眼,使用SPSS11.5统计软件包的卡方检验及Spearman非参数两两相关分析计算各组数据的差异与相关性。结果Worth四点法和卡洞法测得优势眼在习惯屈光矫正下差异无显著性(n=61,P>0.05),也无显著相关性(P>0.05);在屈光全矫下两种方法的差异有显著性(n=140,P=0.001),但无显著相关(P>0.05);而卡洞法在两种条件下测量的优势眼眼别差异有显著性(n=61,P<0.0001),并且存在显著相关(r=0.782,P<0.0001);Worth四点法在两种条件下测量的优势眼眼别差异有显著性(n=62,P<0.0001),并且两者显著相关(n=62,r=0.517,P<0.0001)。结论Worth四点法与卡洞法是两种测量优势眼的方法,在习惯屈光矫正下与屈光全矫状态下测得的优势眼差异有显著性。医学验光时采用Worth四点法测量在习惯矫正下的优势眼的方法尚需进一步研究以确定其可靠性。     

自身抗体在原发性胆汁性肝硬化中的应用分析

目的评价自身抗体对原发性胆汁性肝硬化患者诊断的应用价值,探讨该病与自身抗体的关系。方法用欧蒙印迹法、间接免疫荧光法、免疫金标法检测血清AMA-M2,ANA,ENA和ds-DNA,对结果进行统计、分析,与17例PBC、32例非PBC和35例健康对照进行比较。结果PBC组AMA-M2阳性率为94.1%,特异性为100.0%,非PBC组及健康对照组检出率为零。ANA,ENA和Ads-DNA检测结果在PBC组、非PBC肝硬化组、健康对照组分别为58.8%,56.2%和5.7%,前两组与对照组差异有显著性(P<0.01)。ENA检出率,PBC组23.5%、非PBC肝硬化组28.1%、对照组检出率为0%。结论AMA-M2是PBC的重要血清学标志,同时也是该病的重要诊断手段之一。PBC患者的AMA-M2阳性与临床病情无关,ANA,ENA和Ads-DNA对PBC的诊断无特异性帮助。     

医用X线工作人员晶状体检查与分析

目的评估长期小剂量电离辐射对晶状体的影响。方法通过对医用X线工作人员178人(356眼)晶状体的检查,对该组人员的受照剂量与晶状体点状浑浊程度之间的关系进行分析。结果医用X线工作人员晶状体点状浑浊程度随累积受照剂量和年平均受照剂量的增加而增高。结论医用X线工作人员晶状体点状浑浊程度的增高可能与长期接触小剂量X线照射有关。