DNA Ploidy is an Independent Predictor of Survival in Breast Invasive Ductal Carcinoma: A Long-term Multivariate Analysis of 393 Patients

Purpose

To evaluate “classic” prognostic parameters, as well as DNA ploidy and S-phase fraction (SPF), in relation to disease-free (DFS) and disease-specific (DSS) survival in breast invasive ductal carcinoma (IDC) with long-term follow-up study.

Methods

The study involved 393 patients with IDC and median follow-up of 134 months (50–240). Histological grading, tumor size, axillary nodal involvement, pathological staging and hormone receptor status were considered as established prognostic markers. Ploidy and SPF were determined prospectively by DNA flow cytometry using fresh/frozen tissue. A Cox regression model was used for statistical analysis of the prognostic variables.

Results

There were 105 (26.7 %) deaths and 140 (35.6 %) disease recurrences during follow-up. Two hundred thirty-one (58.8 %) tumors were aneuploid. High SPF and aneuploidy were associated with tumors with higher grade of differentiation, greater size and negative hormone receptors. Higher SPF and advanced disease stage are correlated. In univariate analysis, all the clinicopathological and cytometric features, including patients <40 years and a subgroup presenting hypertetraploid/multiploid tumors, are significantly correlated with clinical outcome, apart from SPF and estrogen receptors for DFS. In multivariate analysis, nodal involvement, DNA aneuploidy and lack of progesterone receptors (for DSS) retained statistically significant association with shorter survival. In node-negative patients, ploidy (for DFS) and estrogen receptors (for DSS) significantly predicted survival. In both subgroups of node-positive patients and those (n = 195) with intermediate differentiation tumors (G2), aneuploidy was an indicator of worse prognosis.

Conclusions

Along with nodal status and hormone receptor expression, DNA ploidy is an independent predictor of long-term survival in IDC.     

Rapid Toluidine Blue Stain for Mohs'' Micrographic Surgery

BACKGROUND: Toluidine blue is a useful stain for detecting basal cell carcinoma during Mohs' micrographic surgery. OBJECTIVE: To demonstrate the efficacy of alkalinization on the toluidine blue stain. METHODS: A 1% aqueous toluidine blue-1% aqueous sodium borate solution was used to stain microscope slides for basal cell carcinoma during Mohs' micrographic surgery. RESULTS: Total toluidine blue staining time was reduced to less than 2.5 minutes, without compromising the quality of the stain. CONCLUSIONS: The rapid toluidine blue stain reduces staining time while maintaining staining quality, including the advantages specific to the toluidine blue stain.     

Mitochondrial tRNALeu isoforms in lung carcinoma cybrid cells containing the np 3243 mtDNA mutation

We have investigated the representation of structural isoforms of the two mitochondrial leucyl tRNAs in lung carcinoma cybrid cell lines containing the np 3243 (MELAS) mtDNA mutation, alone or in combination with the np 12300 suppressor mutation. The mutant tRNALeu(UUR) is aminoacylated very poorly or not at all, whereas the suppressor tRNALeu(CUN) is efficiently aminoacylated. Deacylated mitochondrial tRNALeu(CUN) is present, in all human cells tested, in two structural isoforms that are separable on denaturing gels, indicating a difference in primary structure. The ratio of the two isoforms differs between cell types and is strongly biased towards one isoform in lung carcinoma cybrids containing high levels of the np 3243 mutation, compared with control cybrids. We propose that structural modification of tRNALeu(CUN) could be a natural suppression mechanism for the np 3243 and other mitochondrial tRNALeu(UUR) mutations and could underlie some of the phenotypic variability of np 3243 disease.      

Comparison of Mohs Micrographic Surgery and Wide Excision for Extramammary Paget''s Disease

BACKGROUND: Extramammary Paget's disease is a rare cutaneous adenocarcinoma that occurs in an apocrine gland distribution mainly in the anogenital region. OBJECTIVE: To formulate treatment recommendations for this rare disease, we examined clinical and follow-up data of patients with it. METHODS: A retrospective review is given about the treatment and outcome for 95 patients at Mayo Clinic, Rochester, Minnesota, and Scottsdale, Arizona, between 1976 and 2001. The literature regarding diagnosis and treatment of this disease is also reviewed. RESULTS: Of the 95 patients, 86 had primary disease and 9 had recurrent disease. At mean follow-up (wide excision, 65 months; Mohs surgery, 24 months), disease had recurred in 18 of 83 (22%) who underwent standard wide excision, compared with recurrence in 1 of 12 (8%) who had the Mohs micrographic excision. CONCLUSION: Mohs micrographic surgery compares favorably with wide excision. Intraoperative immunostaining with cytokeratin 7 is helpful in delineating disease, as are preoperative scouting biopsies and photodynamic diagnosis.     

New anti-human immunodeficiency virus immunoblot assays resolve nonspecific western blot results

BACKGROUND: Two new anti-human immunodeficiency virus type 1 and 2 (HIV- 1/2) immunoblot assays, which use recombinant and synthetic antigens for antibody detection have become available. These assays might be able to resolve indeterminate anti-HIV Western blot results. STUDY DESIGN AND METHODS: The new anti-HIV immunoblot assays were used to test 67 samples from 31 HIV infected and 24 noninfected persons showing various anti-HIV Western blot patterns. RESULTS: The immunoblots correctly identified eight HIV-2-positive samples and five late-stage HIV-1-positive samples. After 30 samples from 18 seroconverting persons were tested, the Western blot showed reactivity in 24 of 30 samples and the immunoblots showed reactivity in 21 of 30 samples (difference not significant). In 16 of 30 seroconversion samples, both immunoblot assays produced HIV-1-positive results. In accordance with various criteria for interpretation, the number of positive Western blot results was as follows: Centers for Disease Control and Prevention, 16 of 30 and World Health Organization, 13 of 30. When samples from non- HIV-infected persons were tested, both immunoblot assays resolved 20 (83%) of 24 samples showing indeterminate Western blot reactivity. CONCLUSION: Although the sensitivity of the immunoblot assays should be validation in larger studies, the mere specificity of these assays makes them valuable for the exclusion of HIV infection in Western blot- indeterminate, HIV p24 antigen-negative samples.     

认知暴露疗法治疗创伤后应激障碍

目的:探讨认知暴露疗法结合药物治疗创伤后应激障碍(post-traumaticstressdisorder,PTSD)的效果。方法:于2003-06/2004-06选择第四军医大学西京医院心身科、西安交通大学第一医院和西安市精神卫生中心的门诊和住院PTSD患者20例。均符合美国精神疾病诊断与统计手册第四版PTSD诊断标准。将20例患者按随机数字表法分为结合治疗组和药物治疗组,每组10例。一组采用认知暴露疗法结合药物干预,另一组采取单纯药物干预。认知暴露疗法方案:向患者讲解创伤应激的有关知识、呼吸再训练、放松训练、创伤记忆暴露、自我重复、认知治疗。治疗次数固定,共7次心理治疗会谈。药物治疗方案:①苯二氮革类药物:罗拉,0.5～1.0mg,3次/d,口服。②抗抑郁药物,盐酸氟西汀,20mg,1次/d。于治疗前24h内、临床治愈后24h内、治疗结束后第91天进行PTSD症状清单量表(主要包括3个分量表:再体验、回避和高激惹)、症状自评量表(包括9个因子,分别是躯体化、强迫症状、人际关系敏感、抑郁、焦虑、敌对、恐怖、偏执和精神病性,还有一个其他项目,共计为10个因子)、贝克抑郁问卷、状态-特质焦虑问卷、汉密顿抑郁量表、汉密顿焦虑量表评定。各量表评定分数越高症状越严重。结果:随访期间无一例失访。①PTSD症状清单量表总分及其回避、高激惹两个分量表评分变化在两组之间无差异,而两组间创伤性再体验分量表评分差异有显著性意义(F=5.502,P<0.05);量表总分及回避、高激惹两个分量表评分组内比较差异有显著性意义(F=8.912,9.399,6.007,P<0.01),创伤性再体验分量表评分则无显著差异;两两比较结果发现,量表总分在3个测量点间差异均有显著性意义(t=2.342,3.547,3.124,P<0.05),表明治疗后比治疗前症状显著改善,随访时有显著回升,但仍显著好于治疗前;创伤性再体验分量表评分治疗后显著低于治疗前(t=2.804,P<0.05),其余时间点比较差异不显著;回避和高激惹分量表评分治疗后及随访时均显著低于治疗前(t=3.542,3.124,5.246,4.138,P<0.05),治疗后与随访时差异不显著。②症状自评量表总分两组间比较差异不显著,组内比较差异有显著性意义(F=41.21,P<0.01),量表总分的时间变化与分组之间无交互作用。两两比较结果发现,症状自评量表总分治疗后及随访时均显著低于治疗前(t=10.526,6.354,P<0.01),治疗后与随访时差异不显著。③贝克卓艹抑郁问卷、状态-特质焦虑问卷、汉密顿抑郁量表、汉密顿焦虑量表评分两组间均无差异;各量表组内比较差异有显著性意义(F=56.701,33.165,33.222,33.491,P<0.01),量表评分与分组之间有交互作用(F=9.615,7.110,4.380,3.953,P<0.05)。④结合治疗组有2名不再符合PTSD诊断,药物治疗组仍全部符合PTSD诊断,两组比较无显著差异(χ2=2.004,P>0.05)。结论:认知暴露疗法能够弥补药物治疗的效果短和无法进行认知重建的不足,减少创伤性应激障碍患者的复发,有助于PTSD患者的心理康复。     

低氧促进大鼠骨骼肌成肌细胞体外增殖及重金属钴的作用

目的:观察低氧促进大鼠骨骼肌成肌细胞增殖的作用,并分析CoCl2对成肌细胞增殖的影响。方法:实验于2005-05/2006-07在解放军军事医学科学院基础医学研究所神经肌肉发育研究室完成。①低氧CO2温箱(Forma Scientific,美国);CoCl2(北京化工厂)。②选取4～5周龄Wistar雄性大鼠5只,脱颈处死无菌切取后腿肌群,剪除脂肪和筋膜,制备单细胞悬液。以连续贴壁法筛选大鼠骨骼肌成肌细胞,接种于96孔板进行单克隆培养。采用成肌细胞特异性标志抗原desmin免疫化学染色,鉴定成肌细胞标志蛋白——结蛋白的表达,弃去desmin阴性的细胞克隆,继续培养desmin阳性的细胞克隆,隔天换液1次,7d进行酶消化传代,获得大量扩增的细胞,并可冻存复苏,用于实验。③以1×107L-1接种于20个35mm培养皿中,接种细胞3h后,将培养皿随机数字表法分为5组:常氧对照组、轻度低氧组、中度低氧组、CoCl2组、轻度低氧 CoCl2组,4皿/组。常氧对照组置于体积分数为0.2的O2常规氧气环境中;轻、中度低氧组分别于低氧温箱中维持体积分数为0.1与0.03的O2低氧环境中;CoCl2组向培养皿中加入终浓度为15μmol/L的CoCl2;轻度低氧 CoCl2组向细胞培养皿中加入终浓度为15μmol/L的CoCl2后,放入体积分数为0.1的O2低氧温箱。低氧培养24,48,72h时,各组取出培养皿进行细胞消化离心,倒置相差显微镜下观察细胞生长情况,血球计数板计数法进行细胞计数。④以1×108L-1接种于8个60mm培养皿中,接种细胞3h后,将培养皿随机数字表法分为2组:常氧对照组、轻度低氧组,4皿/组。两组干预措施同细胞计数过程。低氧培养48h时,两组取出培养皿进行细胞消化离心,流式细胞仪检测细胞周期。结果:①骨骼肌成肌细胞的单克隆培养结果:成肌细胞可在体外存活6个月,增殖旺盛期约为3个月,可传代15次以上。单克隆培养2周后,可以由单个细胞长满96孔板中的1个孔,并不断扩增,最终可以得到细胞类型专一单克隆化的成肌细胞。②成肌细胞特异性标志抗原desmin鉴定结果:镜下不同视野desmin阳性率达100%,即培养的成肌细胞单克隆纯度达100%。③细胞计数结果:与常氧对照组比较,低氧培养24,48,72h时轻、中度低氧组均可明显促进大鼠骨骼肌成肌细胞体外增殖,且轻度低氧组作用尤为明显(t=4.98,P<0.001);CoCl2组无明显变化,但轻度低氧 CoCl2组细胞数量显著增加(t=4.62,P<0.001)。④细胞周期分布:与常氧对照组比较,低氧48h时轻度低氧组处于S期的细胞明显增多[(26.67±0.89)%,(65.43±0.23)%,t=2.36,P<0.01],且增殖指数亦显著上升(33.4%,67.1%,t=2.15,P<0.01)。结论:①轻中度低氧可以促进大鼠骨骼肌成肌细胞的增殖,为体外大量扩增细胞提供了新思路。②CoCl2对骨骼肌成肌细胞没有促增殖作用。     

Performance of three generations of anti-hepatitis C virus enzyme- linked immunosorbent assays in donors and patients

BACKGROUND: Prevention of posttransfusion non-A,non-B hepatitis in recipients of blood components improved considerably with the introduction of the second-generation of hepatitis C virus (HCV) antibody tests. In 1993, third-generation HCV antibody assays were introduced in Europe. STUDY DESIGN AND METHODS: The performance of three generations of anti-HCV enzyme-linked immunosorbent assay (ELISA) (ELISA-1, -2, -3) was compared in routine blood donor screening (99,394 donations were tested with ELISA-1, 167,999 donations with ELISA-2, and 262,090 donations with ELISA-3) and in serial samples from nine patients with documented acute posttransfusion HCV infection. RESULTS: Eight (0.01%) repeat donors, previously negative in ELISA-1, were found positive in ELISA-2 and were confirmed as positive in second-generation recombinant immunoblot assay and/or cDNA polymerase chain reaction. In the donor population, no difference in the sensitivity of ELISA-2 and - 3 was observed. The specificity of the three generations of ELISAs was comparable (99.8, 99.7, and 99.7%). In seroconversion samples, ELISA-2 and -3 detected HCV antibodies at the same time in seven patients, but in two patients, ELISA-3 found HCV antibodies, respectively, 63 and 77 days earlier than ELISA-2 did. In the seroconversion samples, ELISA-2 and -3 were significantly more sensitive than second- and third- generation recombinant immunoblot assays. CONCLUSION: ELISA-3 did not detect more HCV-infected individuals in a donor population that previously tested negative in ELISA-2, but it did detect HCV antibodies earlier in some patients with acute HCV infection. ELISA-2 and -3 were significantly more sensitive than second- and third-generation recombinant immunoblot assays.