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71.
应用非同位素标记PCR-SSCP技术对34例大肠癌组织p53基因第五外显子突变进行了检测。结果:50%(11/22)结肠癌,8.3%(1/12)直肠癌存在p53基因突变,低分化腺癌高于高、中分化腺癌(P<0.05),DukesB、C期高于DukesA期(P<0.05),p53基因第五外显子突变在已发生淋巴转移的大肠癌组织中呈现较强的趋势。提示:检测结肠癌原发灶p53基因第五外显子突变,对指导结肠癌的诊断、治疗及预后判断具有一定意义 相似文献
72.
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. 相似文献
73.
Willem E. Corver Gert Jan Fleuren Cees J. Cornelisse 《Journal of immunological methods》2002,260(1-2):97-107
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. 相似文献
74.
Objective To investigate the effect of rosiglitazone on p38 mitogen-activated protein kinase (p38MAPK) pathway in polycystic kidney cyst-lining epithelial cells. Methods The cyst-lining epithelial cells (PKD cells) from human polycystic kidney were treated with rosiglitazone (10 μmol/L), peroxisome proliferator-activated receptor-γ (PPARγ) inhibitor GW9662 (10 μmol/L), rosiglitazone (10 μmol/L) +GW9662 (10 μmol/L), p38MAPK specific inhibitor SB203580 (10 μmol/L), SB203580 (10 μmol/L)+ rosiglitazone(10 μmol/L) for 2 hours followed by epidermal growth factor (EGF) stimulation. Protein expressions of p38, phuspho-p38 (p-p38) and proliferating cell nuclear antigen (PCNA) were detected by Western blot. p38 mRNA was examined by RT-PCR. Expression of c-fos and c-jun was observed by immunocytochemistry. Results (1) EGF markedly up-regulated the expressions of p38, p-p38, PCNA, c-fos anti c-jun compared with control group (P<0.01). (2) Compared with EGF treated group, rosiglitazone significantly reduced p38 activation and mRNA expression (P<0.01, respectively). Rosiglitazone, rosiglitazone+SB203580 could significantly down-regulated p-p38, PCNA, c-fos and c-jun expression (P<0.01, respectively) with no significant difference between these two groups. (3) GW9662 partially reversed the reduction effect of rosiglitazone. Conclusions Rosiglitazone can inhibit proliferation of autosomal dominant polycystic kidney disease cyst-lining epithelial cells partially through down-regulating p38 activation and reducing c-fos, c-jun and PCNA expression. The above effect of rosiglitazone is in part PPARγ-independcnt. 相似文献
75.
瘢痕疙瘩成纤维细胞p53基因突变的研究 总被引:5,自引:0,他引:5
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。 相似文献
76.
ΔNp63蛋白在膀胱移行上皮癌中的表达及其临床意义 总被引:3,自引:1,他引:2
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。 相似文献
77.
乳腺癌组织P21WAF1基因多态性与蛋白表达的关系 总被引:2,自引:0,他引:2
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。 相似文献
78.
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。 相似文献
79.
80.