人端粒酶催化亚单位mRNA的实时荧光RT-PCR定量检测

目的 :建立可定量测定人端粒酶催化亚单位 (hTERT)mRNA的实时荧光反转录PCR方法。方法 :用Trizol 试剂从HepG2细胞中分离总RNA ,然后反转录为cDNA。利用一对基因特异引物和一条TaqManMGB探针 ,以实时荧光PCR定量hTERT的cDNA。同时定量测定人 β 肌动蛋白 (hBA)的mRNA作为内对照。用梯度稀释的克隆的人 β 肌动蛋白基因制作标准曲线。结果 :标准曲线的相关系数为 1.0 0。实时荧光反转录PCR方法的平均变异系数为 7.1%。HepG2细胞hTERTmRNA与hBAmRNA的比值为 (1.3± 0 .3)× 10 -4。结论 :建立了可定量测定人端粒酶催化亚单位 (hTERT)mRNA的实时荧光反转录PCR方法。本方法的建立将有助于研究端粒酶的生物功能     

胃肠道间质瘤临床病理及免疫组织化学特征

目的 探讨胃肠道间质瘤的免疫组织化学特征，为其诊断及鉴别诊断和预后提供依据。方法 对消化道内169例间叶源性肿瘤进行免疫组织化学标记和形态学观察，确诊113例胃肠道间质瘤。结果 肿瘤多见于胃，临床常见首发症状为消化道出血及腹部包块。瘤细胞主要有梭形细胞及上皮样细胞两种形态，梭形细胞型70例，上皮样细胞型10例，混合细胞型33例。相对良性33例，交界性26例，恶性54例。免疫表型：CD117阳性112例，CD34阳性102例，阳性率分别为99．1％及90．2％，且呈弥漫强阳性表达。结论 胃肠道间质瘤是消化道最常见间叶源性肿瘤，以胃内多见；主要有2种细胞形态和3种组合形式；确诊需要依靠CD117、CD34等免疫标记物配合。     

RIN1在膀胱尿路上皮癌中的表达及临床特征

目的探讨膀胱尿路上皮癌中RINl蛋白的表达与临床病理学特征的关系。方法用sP免疫组化法检测手术切除的88例原发性膀胱尿路上皮癌标本和20例癌旁正常组织标本。所有组织用10％福尔马林固定。石蜡包埋,连续切片,厚度4μm。所有上述患者术前均未接受放疗,化疗。通过卡方检验,分析RINl表达与膀胱尿路上皮癌的临床病理学特点。结果免疫组化法发现,88例膀胱尿路上皮癌中有49例RINl表达升高,20例癌旁组织的RINl表达正常。RINl的表达与肿瘤的病理分级（P〈0．05）和临床分期（P〈0．05）呈正相关,与年龄、性别、肿瘤大小、是否多发则无相关性（P〉0．05）。结论RINI在膀胱尿路上皮癌中存在高表达,且在膀胱癌中发挥着重要作用。     

Idiopathic multicentric Castleman's disease. A clinicopathologic and immunohistochemical study of five cases

Several lymphoproliferative disorders may be interpreted as multicentric Castleman's disease (MCD) clinicopathologically. These include HIV infection, autoimmune-disease-associated lymphadenopathy, idiopathic plasmacytic lymphadenopathy with polyclonal hyperimmunoglobulinemia, "idiopathic MCD", POEMS syndrome (polyneuropathy, anasarca, organomegaly, endocrinopathy, M-proteins, and skin lesions), and non-Hodgkin's lymphomas. Among these, idiopathic MCD appears to be relatively rare. We report on the clinicopathologic and immunohistologic findings of five cases of idiopathic MCD and discuss the problems regarding their differential diagnosis. Some of the characteristic clinical findings of POEMS syndrome, including hepatosplenomegaly, skin change, endocrine abnormalities and anasarca, were present in all five cases. However, during the course of disease, minimal diagnostic criteria for POEMS syndrome, i.e., monoclonal plasma cell proliferation and sensory motor neuropathys, were absent in all five cases. The serum interleukin-6 level and the vascular endothelial growth factor level were found to be elevated in two of the cases examined. Various autoantibodies were detected in three cases. However, none of them fulfilled the diagnostic criteria for any of the definite autoimmune-disease. Histologically, three lesions exhibited a mixed type of Castleman's disease, and two exhibited the hyaline-vascular type. The majority of the germinal centers were of the hyaline-vascular or epithelioid germinal type, with a few hyperplastic germinal centers. The interfollicular area was characterized by prominent vascularity. Moderate to large sheets of plasma cells were observed in three mixed type cases. The polytypic nature of B-lymphocytes was demonstrated by immunohistochemistry and polymerase chain reaction. Immunohistochemical study demonstrated that the majority of germinal centers exhibited a tight/concentric pattern of FDC network. Few CD57-positive T-cells were observed in the hyaline-vascular and epithelioid follicles. The lack of CD57-positive T-cells appears to be related to the formation of abnormal germinal centers in the MCD.     

Aurora kinase expression in colorectal adenocarcinoma: correlations with clinicopathological features, p16 expression, and telomerase activity

The clinicopathological roles of aurora kinase protein have not been studied in depth in colorectal cancer. The aim of the present study was to investigate the clinicopathological roles of aurora kinase protein expression in a large cohort of patients with colorectal adenocarcinoma with tight methodology and close follow-up. Aurora kinase protein expression was investigated in 200 patients (110 men, 90 women) with colorectal adenocarcinomas by immunohistochemistry. The findings were correlated with the clinicopathological features, p16 expression, and telomerase activity of colorectal adenocarcinomas. Aurora kinase protein was detected in 48.5% (97/200) of patients with colorectal carcinoma. The protein was more frequently detected in patients with well or moderately differentiated colorectal carcinomas than in poorly differentiated colorectal adenocarcinomas (52% versus 31%, P = .004). Mucinous adenocarcinomas were less often positive for the protein (16% versus 56%, P = .0001). Aurora kinase protein expression was more commonly seen in carcinomas in the rectum, sigmoid, and descending colon than in the proximal colon (55% versus 36%, P = .01). Also, aurora kinase protein expression was related to the expression of p16 protein (P = .001) and correlated inversely with the level of telomerase activity in colorectal carcinomas (P = .005). To conclude, aurora kinase protein is expressed in a subset of colorectal carcinoma. The expression of aurora kinase protein was found to be related to the distal location, grade of tumor, p16 expression, and telomerase activity. These findings may be important to select patients to benefit for clinical trials of aurora kinase inhibitors.     

子宫内膜异位症患者端粒酶及端粒酶催化亚单位的表达研究

目的探索研究不同分型的子宫内膜异位症患者体内腹腔液中端粒酶及端粒酶逆转录酶的表达及对子宫内膜异位症发展的影响。方法在腹腔镜下收集64例子宫内膜异位症手术患者体内腹腔液,应用酶联免疫吸附实验（ELISA）及荧光原住杂交实验（FISH）方法分别检验收集的腹腔液中端粒酶的阳性表达及端粒酶逆转录酶（hTERC）的扩增情况,并与对照组（盆腔炎患者）相比较。结果应用ELISA方法检验出子宫内膜异位症组腹腔液中端粒酶阳性率与对照组差异无统计学意义;FISH实验检验出子宫内膜异位症组腹腔液中人类端粒酶逆转录酶扩增阳性率0-Ⅱ期与对照组差异无统计学意义;Ⅲ-Ⅳ期患者于对照组差异有统计学意义。结论子宫内膜异位症患者腹腔液中端粒酶逆转录酶表达均高于实验对照组,其可能与异位子宫内膜的恶变及增值活性较高相关。     

Angiosarcoma of the uterus: report of 2 new cases with deviant clinicopathologic features and review of the literature

A few cases of uterine angiosarcoma have been detailed in the literature: 2 new cases are herein described featuring some unusual clinical or phenotypic differences compared with previously published cases, such as occurrence in premenopausal age or a poorly differentiated histology. The patients were 35 and 81 years old, and both presented with extrauterine spread and evidence of distant metastases. Grossly, they were fleshy, hemorrhagic, and necrotic. Microscopically, they were made up of poorly differentiated, epithelioid, or spindle cells. In one case, the neoplastic growth deceitfully recalled a poorly differentiated leiomyosarcoma showing focal rudimentary endothelial differentiation. The other index case was characterized by a more pronounced vascular pattern, the neoplasm was composed of spindle cells arranged in loose channels. A diffuse immunopositivity for CD31, CD34, and factor VIII–related antigen was detected. Tumor cells were negative for other tested antigens including keratins, desmin, actins, and H-caldesmon. One patient died shortly during the follow-up, whereas the other is alive with evidence of disseminated disease. Occurrence in childbearing age or a deviant histologic pattern, as documented in this report, may be added to the clinicopathologic spectrum of uterine angiosarcoma.     

AML患者端粒酶逆转录酶mRNA的表达与端粒酶活性的检测

目的:建立一种实时荧光RT-PCR方法,定量检测急性髓细胞白血病(AML)患者外周血单个核细胞端粒酶逆转录酶(hTERT)mRNA的表达水平,观察其与AML发生及复发的关系,探讨hTERT mRNA的表达水平与端粒酶活性的相关关系。方法:采用实时荧光RT-PCR与Lightcycler荧光PCR仪定量检测hTERT mRNA的表达水平,采用PCR-ELISA检测端粒酶活性。结果:①AML首治患者、复发患者、完全缓解期患者以及健康体检者NhTERT分别为299.2±292.8、550.1±441.3、14.0±9.2和12.3±6.7,与健康体检者和完全缓解期患者比较,AML首治患者、复发患者hTERT mRNA表达水平显著升高,而且AML复发患者hTERT mRNA的表达水平显著高于首治患者;②AML首治患者、AML复发患者、AML完全缓解期患者以及健康体检者端粒酶的活性分别为32.8%±24.3%4、8.6%±31.4%、7.4%±5.1%和7.6%±3.6%,AML首治患者、复发患者端粒酶活性显著升高,而且AML复发患者端粒酶活性显著高于首治患者;③hTERT mRNA表达水平与端粒酶的活性呈现良好的相关性,相关系数为0.78。结论:hTERT mRNA表达水平与端粒酶活性的上调是AML发生与复发的一个重要因素,且hTERT mRNA表达水平与端粒酶活性呈良好的正相关。