The effect of corona discharge on Der p 1

BACKGROUND: Reduction of house dust mite allergens in the domestic environment can play an important part in reducing sensitization and in the amelioration of symptoms in atopic individuals. Chemical and physical methods have been tried with varied levels of success. The present paper presents a novel electrostatic way of destroying Der p 1, the major mite allergen. OBJECTIVE: To assess the efficacy of negative Trichel, negative continuous glow, positive pulse and positive continuous glow corona in destroying Der p 1. To determine whether ozone has any effect on the integrity of Der p 1 in the experimental conditions present. METHODS: A simple point-to-plane apparatus was used to irradiate samples of Der p 1 for periods of 1, 15, 30, 45, 60, 120, 180, 240 and 300 min. Controls were exposed to the atmosphere with no corona products present for the equivalent time. The effect of the corona by-product ozone was investigated alone by exposing samples of Der p 1 to molecular ozone for 60 min. Der p 1 concentration was quantified by two-site monoclonal antibody ELISA. RESULTS: High current negative glow resulted in a 67.37% reduction in Der p 1 concentration after 300 min compared with a 50.5% reduction from a low current Trichel regime. High current positive glow corona gave a reduction of 25.22% while a low current positive pulse corona caused a 13.72% reduction after 300 min. All these reductions were statistically significant (P < 0.05) compared with unexposed controls. Negative corona always gave greater percentage reductions in Der p 1 concentration for each time exposure investigated. The pattern of percentage reduction follows an exponential rise to maximum relationship in respect to time. Samples of Der p 1 were not affected by exposure to molecular ozone. CONCLUSION: These data indicate corona products to be a powerful new method of destroying Der p 1 allergen that is not dependent on the presence of the oxidizing corona product ozone.     

Software compensation improves the analysis of heterogeneous tumor samples stained for multiparameter DNA flow cytometry

Background: High concentrations of propidium iodide (PI), in combination with fluorescein isothiocyanate (FITC) and R-phycoerythrin (RPE) used for multiparameter DNA flow cytometry (FCM), cause spectral cross-talk into the green fluorescence channel (FL1). We have evaluated the use of post-acquisition software compensation (N-Color Compensation) in order to correct this spectral cross-talk caused by PI. Method: Cell mixtures were prepared consisting of keratin 8/18 FITC labeled, keratin 8/18 RPE labeled, and unlabeled MCF-7 breast carcinoma cells. DNA was stained with PI (100 μM). Post-acquisition software compensation was applied to correct the spectral cross-talk of PI fluorescence. Secondly, the distribution of the Ki-67 (FITC) protein during the cell cycle (PI) of SiHa cervical carcinoma cells (no software compensation) was compared to the Ki-67 expression pattern of SiHa cells, simultaneously stained for keratin 8 (RPE), after applying software compensation. Finally, software compensation was used to compare the relative levels of PCNA and p53 expression in two clinical ovarian cancer ascites specimens, stained for PCNA or p53 (FITC), keratin 8/18 (RPE), and DNA (PI), with a known p53 status (positive and negative, respectively). Results: The Ki-67 cell cycle-dependent pattern of a triply stained sample (Ki-67 (FITC), keratin 8 (RPE), and DNA (PI)) is restored after software compensation and the results are comparable to the Ki-67 distribution of a sample stained solely for Ki-67 and DNA. P53 expression could only be resolved after using software compensation in the p53 positive ovarian ascites (OA) sample. Conclusions: We conclude that software compensation is a robust and reliable post-acquisition method for the correction of RPE/PI spectral cross-talk, permitting better identification of weakly expressed proteins in heterogeneous clinical tumor samples stained for multiple cellular antigens and DNA using PI.     

长波紫外线照射对小鼠黑素瘤细胞的影响

[目的]研究不同剂量UVA照射对体外培养B_16小鼠黑素瘤细胞的形态、增殖率、黑素含量和酪氨酸酶活性的影响。[方法]选用B_16小鼠黑素瘤细胞 ,经不同剂量的UVA照射后 ,采用MTT法测定细胞的增殖 ,参照HideyaAndo等的方法测定酪氨酸酶活性和黑素含量。[结果]UVA照射后 ,黑素含量增加 (P<0.01) ,酪氨酸酶活性提高 (P<0.05) ,细胞增殖率上升 ,照射剂量在0.6、1.2、1.8J/cm2时 ,细胞增殖率的变化有显著性 (t=2.495、2.645、3.081,均为P<0.05)。[结论]随着照射剂量的增加 ,黑素含量增加 ,酪氨酸酶活性提高 ,细胞的增殖率增加 ,但UVA的促黑素瘤细胞增殖作用是有限的     

瘢痕疙瘩成纤维细胞p53基因突变的研究

目的　探讨瘢痕疙瘩成纤维细胞中 p5 3基因第 4～ 8外显子的突变规律及其意义。　方法　取瘢痕患者手术切除的瘢痕疙瘩和增生性瘢痕标本各 12例 ,并设患者自身正常皮肤标本及血标本为对照。体外分离、培养上述组织标本的成纤维细胞。采用聚合酶链式反应 单链构象多态性(PCR SSCP)分析方法和基因测序法 ,检测各种组织成纤维细胞中p5 3基因的突变情况。　 结果　 12例瘢痕疙瘩标本中有 9例p5 3基因外显子 4、5、6、7出现点突变和移码突变 ,增生性瘢痕标本、正常皮肤标本及血标本中均未检出突变。　结论　p5 3基因突变是瘢痕疙瘩形成和发展的重要因素之一。     

ΔNp63蛋白在膀胱移行上皮癌中的表达及其临床意义

目的 :探讨 p5 3基因家族新成员截短型p6 3(△Np6 3)在膀胱癌组织中的表达及其意义。 方法 :采用免疫组织化学SP法检测 4 0例膀胱移行上皮癌 (TCC)、6例膀胱内翻性乳头状瘤和 8例正常膀胱移行上皮中△Np6 3的表达 ,并分析△Np6 3表达与膀胱癌病理类型、临床分期的关系。 结果 :正常膀胱移行上皮、膀胱内翻性乳头状瘤、TCC中△Np6 3的阳性表达率分别为 37.5 % (3/ 8)、6 6 .7% (4/ 6 )、10 0 % (40 / 4 0 ) ,组间差异有统计学意义 (P <0 .0 1)。TCCG3 级与G2 级△Np6 3的强阳性、中度阳性表达率显著高于G1级 (P <0 .0 1)。Ta～T1期以△Np6 3弱阳性为主 (6 6 .7% ) ,随TCC浸润程度的增加 ,△Np6 3染色强度逐渐增强。T2 期△Np6 3强阳性表达率为 35 .3% ,T3 ～T4期增至 6 3.6 %。结论 :△Np6 3在TCC中高表达 ,与TCC病理分级、临床分期密切相关 ;△Np6 3可能参与TCC的发生、发展 ,是评估TCC预后的潜在因素之一。     

人乳头瘤病毒多肽与人免疫球蛋白G融合表达

目的 将人乳头瘤病毒16型(Human papillomavirus type 16，HPV-16)的晚期表达蛋白E7上的抗原24肽(从第38位氨基酸到第61位氨基6病毒感染防治酸)与人免疫球蛋白G的重链恒定区融合表达，并以此融合蛋白作为抗原，可能为HPV-1提供免疫治疗方法。方法 利用PCR方法分别扩增HPV-16 E7(38-61)24肽的DNA片段和人免疫球蛋白G的重链恒定区DNA片段，并构建到pEV21a表达载体上，转化入E．coli中表达，利用SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质免疫印迹(Western-blotting)的方法对表达结果进行鉴定。结果 构建的表达载体HPV16E7e／hIgGHCCR-pET21a经酶切鉴定和测序显示序列正确；通过SDS-PAGE和Western-blotting的鉴定，重组融合蛋白Mr约40000，表达量可占菌体蛋白的20％左右。结论 成功构建HPV16-E7的抗原多肽片段和人免疫球蛋白G的重链恒定区的融合蛋白，并可在E．coli中高效表达。     

乳腺癌组织P21WAF1基因多态性与蛋白表达的关系

目的 :探讨乳腺癌组织p2 1WAF1DNA多态性与蛋白表达的关系。方法 :分别采用聚合酶链反应单链构象多态性技术 (PCR -SSCP)和免疫组化S -P法检测 10 0例乳腺癌石蜡包埋组织和 4 0例乳腺良性病变组织p2 1WAF1DNA多态性和蛋白表达。结果 :18% (18/ 10 0 )的乳腺癌组织和 5 % (2 / 4 0 )对照组组织分别出现两种p2 1WAF1DNA多态性 ,前者明显高于后者 ,组间有显著差异 (χ2 =3 94 ,P <0 0 5 ) ;5 0 % (5 0 / 10 0 )的乳腺癌组织和 12 5 % (5 / 4 0 )对照组组织表达p2 1WAF1蛋白 ,前者明显高于后者 ,组间有显著差异 (χ2 =16 84 ,P <0 0 1) ;10 0 % (18/ 18例 )的具有p2 1WAF1DNA多态性的乳腺癌组织p2 1WAF1蛋白表达阳性 ,39% (32 / 82 )的无p2 1WAF1DNA多态性乳腺癌组织蛋白表达阳性 ,p2 1WAF1DNA多态性组明显高于无多态性组 ,组间有显著差异 (χ2 =2 1 95 ,P <0 0 1) ;p2 1WAF1DNA多态性与p2 1WAF1蛋白表达呈正相关 (r =0 5 76 ,P <0 0 1)。结论 :乳腺癌组织p2 1WAF1DNA多态性可能产生不同的转录本并生成相应的蛋白质分子。     

D16S80和D16S125两位点RFLP在多囊肾症状前诊断中的应用

给出了用“混合型”雅可比级数的泰勒平均逼近黎普希兹函数类的点态估计。     

联合检测自发凋亡率及p27kip1在膀胱移行细胞癌中的意义

目的探讨自发凋亡率、p27kip1在人膀胱移行细胞癌中表达的临床意义。方法采用TUNEL法检测自发凋亡细胞,用S’P法检测p27kip1基因在50例膀胱移行细胞癌中的表达,分析各检测指标在上述标本中的表达及与膀胱移行细胞癌分级、临床分期间的关系。结果在膀胱移行细胞癌中,自发凋亡率AI均数为(3.0±1.5)%,p27kip1基因的阳性表达率为58.0%(29/50)。其二者的阳性表达均随膀胱癌分级、分期的升高而降低。p27kip1蛋白阳性组中AI均数显著高于p27kip1蛋白阴性组(P<0.05)。结论p27kip1蛋白和自发凋亡率与肿瘤的恶性程度和进展密切相关,联合检测p27kip1和自发凋亡率有助于更准确地解释和描述膀胱癌的生物学行为。     

p53基因与肝癌

p53肿瘤抑制基因突变与多数恶性肿瘤的发生发展有关,包括肝细胞癌(hepatocelluar carcinoma,HCC)在内的人类恶性肿瘤中至少有50%发生了p53基因改变。因此,以正常p53基因治疗肿瘤就成了研究热点,随着介入放射学(inter-ventional radiology)向纵深发展,经介入放射方法进行肝癌的基因治疗令人关注。本文介绍了p53基因的结构与功能,其与肝癌的病理联系以及在肝癌治疗中的应用。