肝癌发生不同病理阶段血清蛋白质组的比较

目的 比较肝癌发生不同病理阶段血清蛋白质组的表达变化情况,从中寻找特异性标志物.方法 用双向凝胶电泳分离正常人,慢性肝炎、肝硬化和肝癌患者的血清蛋白质,结合质谱技术对差异点进行鉴定; Western blot检测结合珠蛋白β链以验证蛋白质水平的表达.计量资料数据以均数±标准差(x±s)表示,采用方差分析和LSD检验进行两两比较.结果 4组血清的双向电泳图谱经软件匹配分析,并与基质辅助激光解析电离飞行时间质谱相结合,成功鉴定出结合珠蛋白,SAA1、SP40等30个差异蛋白质点,K cluster聚类分析不同的变化模式.在差异蛋白质点中,7个蛋白质点均为结合珠蛋白,呈现由正常→肝炎→肝硬化持续下调、到肝癌又上调的模式.Western blot证实结合珠蛋白β链的表达与双向电泳表达相一致.结论 在肝癌发生的动态过程中各疾病阶段的血清蛋白质差异表达有4种明显变化模式,可能为肝癌的早期诊断和预后提供依据,与肝癌发生发展相关的结合珠蛋白很可能是肝硬化发展到肝癌的一个潜在早期诊断标志物.     

低剂量T-2毒素与脱氧雪腐镰刀菌烯醇单独及联合染毒对小鼠末梢血有核细胞DNA损伤的观察

目的 观察低剂量T-2毒素和脱氧雪腐镰刀菌烯醇(DON)对小鼠末梢血有核细胞DNA损伤的特点及其程度,探索亚临床剂量下的毒性作用.方法 3周龄雌性Balb/c小鼠80只,体质量(14.0±1.5)g.将小鼠按体质量随机分为对照组、T-2毒素组、DON组、T-2+DON联合组,每组20只.经腹腔注射感染毒素,染毒剂量:T-2毒素5 μg·kg-1·d-1,DON 20 μg·kg-1·d-1;对照组给予等量生理盐水.染毒至16周和21周,分别取小鼠尾血,用单细胞凝胶电泳法(SCGE)观察小鼠末梢血有核细胞拖尾率、尾DNA含量、尾长、尾动量变化.结果 ①T-2毒素组,尾DNA含量、尾长、尾动量[16周:(27.71±15.85)%、(13.67±5.56)μm、4.26±3.83;21周:(28.38±15.57)%、(13.83±5.47)μm、4.37±3.82]均增加,与对照组[16周:(11.87±4.61)%、(10.59±6.70)μm、1.34±0.98;21周:(11.31±3.94)%、(10.83±7.05)μm、1.29±1.01]比较,差异有统计学意义(P均<0.05);②DON组拖尾率、尾DNA含量、尾动量[16周:5.62%、(28.13±13.31)%、3.39±2.35;21周:7.71%、(29.17±15.12)%、5.70±4.17]均增加,其中仅拖尾率与对照组(16周:4.34%;21周:4.38%)比较,差异有统计学意义(P均<0.05);③T-2+DON联合组拖尾率、尾DNA含量、尾长、尾动量[16周:6.21%、(30.14±15.48)%、(16.93±6.58)μm、5.54±4.22;21周:8.17%、(30.85±15.76)%、(17.21±6.45)μm、5.70±4.17]均增加,与对照组比较差异有统计学意义(P均<0.05);与T-2毒素或DON组比较,尾DNA含量和尾长均增加(P均<0.05).结论 低剂量T-2毒素和DON均可对小鼠未梢血有核细胞DNA产生明确损伤;T-2和DON联合染毒较单独染毒的损伤作用更明显.     

苦参对阴道毛滴虫蛋白质差异表达的影响

目的寻找阴道毛滴虫的特异蛋白质并探讨苦参对其差异蛋白质表达的影响。方法用固相pH梯度双向凝胶电泳分离阴道毛滴虫总蛋白质,凝胶用银染显色,PDQuest7.4.0软件分析。结果同组实验重复3次以上,其中正常对照组找到蛋白质点平均（435±17）个,匹配率为92.5%;苦参组找到蛋白质点平均（336±26）个,匹配率为85.6%。正常组和苦参组共有96个表达差异点,其中苦参组和正常组比较,只在正常组中表达的点有46个,只在苦参组中表达的点有27个,表达增加〉2倍的点有9个,表达降低〉0.5倍的点有14个。结论建立了阴道毛滴虫正常组和苦参组的双向凝胶电泳图谱,识别了96个差异表达的蛋白质。苦参杀灭阴道毛滴虫的作用可能与上述蛋白质的改变有关。     

The prevalence of microsatellite instability in head and neck squamous cell carcinoma

Purpose  The present study aimed to use the most definitive available techniques to resolve controversy in the literature as to the prevalence of microsatellite instability (MSI) in head and neck squamous cell carcinoma (HNSCC). Methods  Eighty patients with advanced HNSCC were enrolled in the study that examined 20 microsatellite markers with automatic fragment analysis. These markers included ones derived from the NCI reference panel and ones previously reported to detect MSI in HNSCC (HNSCC panel). Results  Only one of 80 tumors could be considered positive for MSI. For this case, both panels showed MSI-High (8/10 positive markers for the NCI reference panel and 6/10 positive markers for the HNSCC panel). Qualitatively, the observed MSI could be classified as Type B MSI. Conclusions  The present results indicate that MSI has a low prevalence in HNSCC.     

二维有限元法分析THA术前试模的重要性

目的：通过全髋关节置换术（THA）术前试模与否与术后有限元模型的建立及力学分析,探讨术前试模的重要性。方法：选取行单侧THA的24例患者作为研究对象,将其分为两组（试模组和对照组）,行THA,术后对患髋摄X线正位片,其原始数据输入有限元软件建立二维有限元模型,并进行力学分析、绘制应力曲线,与正常股骨近端的应力曲线进行比较。结果：试模组的股骨近端生物力学分析优于对照组,其应力曲线图较对照组更接近正常。结论：THA术前精确的试模能够使术后的股骨近端应力分布更接近正常。     

The Remarkable Stability of Chimeric,Sialic Acid-derived α/δ-Peptides in Human Blood Plasma

Peptides are labile toward proteolytic enzymes, and structural modifications are often required to prolong their metabolic half-life and increase resistance. One modification is the incorporation of non-α-amino acids into the peptide to deter recognition by hydrolytic enzymes. We previously reported the synthesis of chimeric α/δ-peptides from glutamic acids (Glu) and the sialic acid derivative Neu2en. Conformational analyses revealed these constructs adopt secondary structures in water and may serve as conformational surrogates of polysialic acid. Polysialic acid is a tumor-associated polysaccharide and is correlated with cancer metastasis. Soluble polysialic acid is rapidly cleared from the blood limiting its potential for vaccine development. One motivation in developing structural surrogates of polysialic acid was to create constructs with increased bioavailability. Here, we report plasma stability profiles of Glu/Neu2en α/δ-peptides. DOTA was conjugated at the peptide N-termini by solid phase peptide synthesis, radiolabeled with ¹¹¹In, incubated in human blood plasma at 37 °C, and their degradation patterns monitored by cellulose acetate electrophoresis and radioactivity counting. Results indicate that these peptides exhibit a long half-life that is two- to three-orders of magnitude higher than natural α-peptides. These findings provide a viable platform for the synthesis of plasma stable, sialic acid-derived peptides that may find pharmaceutical application.     

Simultaneous separation and confirmation of amphetamine and related drugs in equine plasma by non-aqueous capillary-electrophoresis-tandem mass spectrometry

A non-aqueous capillary electrophoresis-mass spectrometry (NACE-MS) method was developed for simultaneous separation and identification of 12 amphetamine and related compounds in equine plasma. Analytes were recovered from plasma by liquid-liquid extraction using methyl tertiary butyl ether (MTBE). A bare fused-silica capillary was used for separation of the analytes. Addition of sheath liquid to the capillary effluent allowed the detection of the analytes by positive electrospray ionization mass spectrometry using full scan data acquisition. The limit of detection (LOD) for the target analytes was 10-200 ng/mL and that of confirmation (LOC) was 50-1000 ng/mL in equine plasma. Capillary electrophoresis (CE) and mass spectrometry (MS) parameters were optimized for full CE separation and MS detection of the analytes. Separation buffer comprised 25 mM ammonium formate in acetonitrile/methanol (20: 80, v/v) plus 1 M formic acid. Sheath liquid was isopropanol-water-formic acid (50:50:0.5, v/v/v). Samples were hydrodynamically injected and separated at 25 kV. Analytes were electrokinetically separated and mass spectrometrically identified and confirmed. This simple, fast, inexpensive and reproducible method was successfully applied to post race equine plasma and research samples in screening for amphetamine and related drugs.     

多发性骨髓瘤117例血清免疫学特征分析

目的分析多发性骨髓瘤（MM）血清免疫学特征。方法对117例MM患者血清进行血清蛋白电泳、免疫固定电泳、免疫球蛋白（IgG、IgA、IgM）及总蛋白,白蛋白定量分析。结果117例MM患者血清蛋白电泳显示M带67例（57．3％）,主要位于1区。免疫固定电泳分型：IgG型62例（53．0％）,IgA型19例（16．3％）,IgM型12例（10．1％）,DL型2例（1．7％）,轻链型22例（18．8％）。同型免疫球蛋白含量显著升高并常伴有非对应组分的偏低。血清总蛋白有不同程度的升高和白蛋白一定程度的降低。结论血清免疫学特征综合分析,对MM的诊断分型和临床分期及预后判断具有重要意义。     

Identification of metabolic pattern and bioactive form of resveratrol in human medulloblastoma cells

Cancer preventive reagent trans-resveratrol is intracellularly biotransformed to different metabolites. However, it is still unclear whether trans-resveratrol exerts its biological effects directly or through its metabolite(s). This issue was addressed here by identifying the metabolic pattern and the bioactive form of resveratrol in a resveratrol-sensitive human medulloblastoma cell line, UW228-3. The cell lysates and condition media of UW228-3 cells with or without 100 μM resveratrol treatment were analyzed by HPLC and LC/MS which revealed (1) that resveratrol was chemically unstable and the spontaneous generation of cis-resveratrol reduced resveratrol's anti-medulloblastoma efficacy and (2) that resveratrol monosulfate was the major metabolite of the cells. To identify the bioactive form of resveratrol, a mixture-containing approximately half fraction of resveratrol monosulfate was prepared by incubating trans-resveratrol with freshly prepared rat brain lysates. Medulloblastoma cells treated by 100 μM of this mixture showed attenuated cell crisis. The overall levels of the three brain-associated sulfotransferases (SULT1A1, 1C2 and 4A1) were low in medulloblastoma cells in vivo and in vitro in comparison with that in human noncancerous and rat normal cerebella; resveratrol could more or less up-regulate the production of these enzymes in UW228-3 cells but their overall level was still lower than that in normal cerebellum tissue. Our study thus demonstrated for the first time that trans-resveratrol is the bioactive form in medulloblastoma cells in which the expression of brain-associated SULTs was down-regulated, resulting in the increased intracellular bioavailability and anti-medulloblastoma efficacy of trans-resveratrol.