采用蛋白指纹图谱技术筛选大肠癌肝转移特异性相关蛋白

目的:应用表面增强激光解析电离飞行时间质谱技术从大肠癌及大肠癌肝转移患者中筛选出大肠癌肝转移患者血清特异性相关蛋白。方法:实验于2005-07/2006-09分别在南方医院消化中心实验室与解放军第一五○医院实验室完成。应用美国CipherGen公司IMAC3(ImmobilizedMetalAffinityCapture,金属亲和表面)芯片和蛋白芯片仪检测44例大肠癌患者及36例大肠癌肝转移患者血清中的蛋白质相对含量。设定所有血清样本检测的蛋白质相对分子质量区间在1500～20000。利用PBSⅡ型蛋白质芯片阅读仪对IMAC3芯片进行检测,所得到的蛋白质以波谱的形式表示。采用BiomarkerWizard软件对2组血清在相同质荷比的蛋白质含量数据进行方差分析,将分析所得到的含量有显著性差异(P<0.05)的蛋白质建立数据库,导入BiomarkerPattern智能统计分析软件,选择相应条件,对其进行分组统计,从而得到能够正确分组的特异性蛋白标志物并构建大肠癌肝转移的诊断模型。采用酶联免疫法检测相同血清标本中的CEA水平,与构建的诊断模型在大肠癌肝转移诊断中作比较。结果:①44例大肠癌患者与36例大肠癌肝转移患者的血清蛋白质在质荷比为2685.64～11813间有16个蛋白质含量有显著差异。②大肠癌组在质荷比为5909处的蛋白质的相对含量高于大肠癌肝转移组[(30.1±9.6)%,(14.5±10.4)%,P≤0.01]。③其中44例大肠癌患者中有38例患者被正确分组,36例大肠癌肝转移患者被正确识别,准确率为92.5%(74/80),灵敏度和特异性分别为100%(36/36),86.4%(38/44)。结论:表面增强激光解析电离飞行时间质谱技术快速、准确、灵敏度、特异性高,通过蛋白芯片仪发现的特异性相关蛋白,有望成为大肠癌肝转移诊断中有应用价值的临床检测指标。     

Clinical value of monitoring eosinophil activity in asthma

The prevalence and clinical features of migraine headache and abdominal migraine were studied in the well defined population of Aberdeen schoolchildren. Ten per cent of all children (2165) aged 5-15 years were given a questionnaire inquiring, among other symptoms, about the history of headache and abdominal pain over the past year. A total of 1754 children (81%) responded. Children with at least two episodes of severe headache and/or sever abdominal pain, attributed by the parents either to unknown causes or to migraine, were invited to attend for clinical interview and examination. After interview, 159 children fulfilled the International Headache Society's criteria for the diagnosis of migraine and 58 children had abdominal migraine giving estimated prevalence rates of 10.6% and 4.1% respectively. Children with abdominal migraine had demographic and social characteristics similar to those of children with migraine. They also had similar patterns of associated recurrent painful conditions, trigger and relieving factors, and associated symptoms during attacks. The similarities between the two conditions are so close as to suggest that they have a common pathogenesis.     

Platelet-derived growth factor promotes human peripheral monocyte activation

Like in the polymorphonuclear leukocyte (PMN), the platelet-derived growth factor (PDGF) purified to homogeneity is capable of inducing monocyte activation responses as evaluated by generation of superoxide anion (O-.2) from membrane-associated oxidase system, release of granule enzymes, and enhanced cell adherence and cell aggregation. Superoxide anion release was maximized at 10 ng/mL PDGF and was comparable to that induced by 10(-7) mol/L formyl-methionyl-leucyl- phenylalanine. The potency of PDGF to induce this response in monocytes was of the same magnitude as that observed in PMNs. Similarly, lysozyme release and monocyte adherence were also increased in a dose-dependent manner and achieved maximal responses at 40 ng/mL concentration of PDGF. The PDGF concentration required to achieve maximal monocyte aggregation was two-fold (60 ng/mL) of that found for PMNs. In contrast to PMNs, a positive correlation (gamma = .93; P less than .01) was observed between the increases of PDGF concentration and beta- glucuronidase release. These findings indicate that PDGF can induce the full sequence of cell activation events in human monocytes similar to human PMNs.     

高效液相色谱法测定尿中美芬妥英及代谢产物4'-羟基美芬妥英

A simple, sensitive and reproducible HPLC assay is described for the determination of mephenytoin and 4'-hydroxymephenytoin in human urine. Phenobarbital was used as an internal standard.The compounds were separated on a U-Bondapack RP-C₁₈ column using a mobile phase of and the UV detectou was set at 210 nm. Calibration curves in the range 0.05～1.00ug/ml for mephenytoin and 0.5～100.0ug/ml for 4'-hydroxymephenytoin were linear (r=0.9998and r=0.9992,respectivesy). The average recovery was 95.10±2。95%,and the relative standard devia- tion within day and day to day was less than 10%。The detection limit for mephenytoin was 25mg/ml and 4‘-hydroxymephenytoin was 50mg。ml。The method was used to study the metabolism of S-mephenytoin 4'-hydroxylatoin in 10healthy volunteers.The 12 h urinary metabolic ratio (MR)and hydroxylation index(HI)were calculated to express interindividual variation in metabolism. Two of them exhibited defective 4'-hydroxylation of S-mephenytoin as poor metabllizers (HI:1349.18 and 409.57;MR:105.29 and 8.25).In the remaining 8 subjects, the ranged from 1.68 to 6.71 and the MR ranged from 0.002 to 0.014,as extensive metabolizers of S-mephenytoin.     

Lack of evidence of neurotoxicity following 8 weeks of treatment with tripotassium dicitrato bismuthate

Objective: To search for evidence of subclinical neurotoxicity in patients treated with tripotassium dicitrato bismuthate. Design: Prospective, controlled, triplicate study using urinary bismuth concentration, magnetic resonance imaging (MRI), nerve conduction studies, visual evoked response and a battery of 10 neuropsychological screening tests. Setting: Out-patient clinics, Walsgrave Hospital, Coventry, UK. Subjects: Fourteen dyspeptic patients; 8 (treatment group) treated with tripotassium dicitrato bismuthate one tablet q.d.s and 6 (control group) treated with ranitidine 150 mg b.d. for 8 weeks. Main outcome measures: Changes in urinary bismuth, MRI, nerve conduction studies, visual evoked response, and neuropsychological tests performed before, immediately after and 8 weeks after the cessation of treatment. Results: In the treatment group the median (range) urinary bismuth concentration was 1 (1–12) ng/ml before treatment, increased to 560 (140–1300) immediately after treatment (P < 0.01, Wilcoxon Rank Sum test) and was still significantly elevated (23 (7–53) ng/ml) 8 weeks after the cessation of treatment. In the patient who recorded the highest urinary bismuth, a high intensity signal appeared in the globus pallidus immediately after treatment and was still present (though diminished in intensity) 8 weeks after the cessation of treatment. This isolated MRI finding was not associated with evidence of subclinical neurotoxicity. No changes in the MRI, nerve conduction studies, visual evoked response and neuropsychological tests were observed among the other patients studied. Conclusions: Bismuth accumulation occurs in patients receiving a conventional course of treatment with tripotassium dicitrato bismuthate but this is not associated with significant changes in the nervous system.     

A clinical study of 77 patients with mucopolysaccharidosis type II

AIM: This study aims to assess the clinical features of 77 South American patients (73 Brazilian) with mucopolysaccharidosis type II (MPS II). METHODS: Details of the patients and their disease manifestations were obtained from a review of medical records, interviews with the patients and/or their families, and physical examination of the patients. RESULTS: Mean birth weight was 3360 g, median age at onset of symptoms was 18 months and median age at diagnosis was 6 years. For the whole sample (median age, 8.2 years; range, 2.8-53.0 years), neurological degeneration, typical pebbly skin lesions, seizures and extensive dermal melanocytosis were found in 23.3, 13.0, 13.0 and 1.3% of the cases, respectively. The most frequently reported echocardiogram abnormality was mitral valve regurgitation. Refraction errors were the most common ophthalmological manifestation. The following characteristics were found to be associated with the severe form of MPS II: earlier age at biochemical diagnosis, higher levels of urinary glycosaminoglycans, language development delay, behavioural disturbances, poor school performance and mental retardation. CONCLUSION: Our results suggest that there is a considerable delay between the onset of signs and symptoms and the diagnosis of MPS II in Brazil (and probably in South America as well), and that many complications of this disease are underdiagnosed and undertreated. Therefore, the implementation of programmes aiming to increase the awareness of the disease, the availability of biochemical diagnostic tests and the provision of better support to affected patients is urgently needed.     

链球菌蛋白对人肝癌Bel-7402细胞的增殖抑制作用及其分子机制

目的:从分子水平观察链球菌蛋白质对人肝癌Bel-7402细胞的生长增殖的抑制作用。方法:实验于2005-10/2006-11在首都医科大学病原生物学系实验室完成。①链球菌32080株购自中国医学细菌保藏管理中心,人肝癌Bel-7402细胞株由首都医科大学细胞生物学系惠赠。②常规培养链球菌并按照Winters方法提取细菌蛋白。③用0,10,20,100mg/L细菌蛋白作用于单层培养的Bel-7402细胞24,48,72h,采用MTT法检测链球菌蛋白对细胞的增殖抑制作用。④Bel-7402细胞经10mg/L或50mg/L细菌蛋白分别作用48,72h后,流式细胞术检测细菌蛋白对癌细胞生长周期时相分布。⑤Bel-7402细胞经50mg/L的细菌蛋白分别作用24,48,72h后,Annexin-v PI双染法检测Bel-7402细胞凋亡率。⑥用50mg/L的链球菌蛋白分别作用于Bel-7402细胞0,24,48,72h后,Western Blot检测链球菌蛋白对Bel-7402细胞凋亡相关蛋白的影响。结果:①链球菌蛋白对Bel-7402细胞的抑制作用:链球菌蛋白能显著抑制人肝癌Bel-7402细胞增殖,100mg/L细菌蛋白作用于细胞24,48,72h的抑制率分别为15.6%,35.6%,50%,呈剂量和时间依赖关系。②细菌蛋白对肝癌细胞生长周期时相分布及细胞凋亡率的影响:经链球菌蛋白(10,50mg/L)作用后,G2/M期Bel-7402细胞比率较正常对照组增加,差异有显著性(P<0.05);流式细胞术检测显示,经50mg/L链球菌蛋白作用后,实验组Bel-7402细胞凋亡率明显高于对照组细胞(P<0.01)。③Bel-7402细胞凋亡相关蛋白的变化:Bel-7402细胞经50mg/L链球菌蛋白分别作用24,48,72h后,细胞色素C的释放逐渐增多,Procaspase-3及Bcl-2蛋白水平降低。结论:链球菌32080株蛋白对人肝癌Bel-7402细胞的生长抑制作用存在着剂量和时间依赖性,并能将细胞的生长周期阻滞于G2/M期,诱导细胞凋亡,影响Bcl-2,procaspase-3和细胞色素C等重要的凋亡相关蛋白水平。