肝细胞癌nm23—HImRNA表达水平及其临床意义

探讨肝细胞癌ｎｍ２３┐ＨＩｍＲＮＡ表达水平与临床病理特征的联系。方法通过Ｎｏｒｔｈｅｒｎｂｌｏｔ杂交技术检测５６个外科手术切除的肝细胞癌及癌旁肝组织中ｎｍ２３┐ＨＩｍＲＮＡ水平，并分析其表达与肿瘤大小、血清ＡＦＰ值、肝内转移或门静脉癌栓、包膜侵犯、ＴＮＭ分期等的关系。结果肝癌组织ｎｍ２３┐ＨＩｍＲＮＡ表达水平与肝内转移和／或门静脉形成癌栓，以及ＴＮＭ分期呈负相关，在ｍＲＮＡ低表达组，６６．７％的肝癌伴肝内转移和／或门静脉癌栓，７３．３％为ＴＮＭⅢ、Ⅳ期肝癌。结论ｎｍ２３┐ＨＩ基因低表达在肝癌浸润过程中具有一定作用，ｎｍ２３┐ＨＩｍＲＮＡ表达水平下降可作为肝内转移的标志。     

hMAM mRNA在乳腺癌组织中的表达

目的：探讨乳腺组织特异性基因ｈＭＡＭ ｍＲＮＡ作为检测乳腺癌微小转移指标的可能性。方法：采用ＲＴ－ＰＣＲ方法,检测ｈＭＡＭ ｍＲＮＡ在乳腺癌组织和相应癌旁正常乳腺组织４４例及胃癌、大肠癌、食管癌、肺癌、卵巢癌组织各５例中的表达。分析ｈＭＡＭｍＲＮＡ表达与乳腺癌临床病理特点之间的关系。实验结果进行统计学处理。结果：４４例乳腺癌组织和相应癌旁组织中均表达ｈＭＡＭ ｍＲＮＡ,其中有５６．８２％（２５／４     

p16基因mRNA及其编码蛋白在人乳腺癌中的表达

目的：研究人乳腺癌中ｐ１６基因ｍＲＮＡ及其编码蛋白的表达水平与肿瘤发生中预后的相关性。方法：采用免疫组化和原位杂交方法分别对１１例乳腺良性肿瘤和５９例乳腺癌中的ｐ１６基因及其编码蛋白进行检测。结果：ｐ１６基因蛋白表达水平与乳腺肿瘤的良、恶性显著相关（Ｐ〈０．０５）;ｐ１６基因ｍＲＮＡ及其蛋白表达水平与乳腺癌的腋窝淋巴结转移呈现显著负相关（Ｐ〈０．０５）,而与乳腺癌的术后自下而上期呈显著正相关（Ｐ〈     

乳腺癌患者辅助化疗前后外周血中ＰＩＰｍＲＮＡ表达变化的研究

目的：通过检测乳腺癌患者辅助化疗前后外周血中泌乳素诱导蛋白（ＰＩＰ）ｍＲＮＡ的表达变化,探讨术后辅助化疗对乳腺癌血液微转移的影响。方法：收集沈阳军区总医院肿瘤科２００６年７月～２００７年９月经术后病理证实的乳腺癌患者５０例,在化疗前及接受２～３周期辅助化疗后应用巢式ＲＴ-ＰＣＲ方法检测这些乳腺癌患者外周血中ＰＩＰｍＲＮＡ的表达。另取乳腺纤维腺瘤１０例、健康志愿者１０例、胃癌５例、结直肠癌５例、食管癌５例、肺癌５例、卵巢癌５例作阴性对照。结果：５０例乳腺癌患者外周血标本中,化疗前１７例ＰＩＰｍＲＮＡ阳性的患者,化疗２～３周期后１３例ＰＩＰｍＲＮＡ转为阴性,转阴率７６.５％（１３／１７）,化疗前ＰＩＰｍＲＮＡ表达阴性的患者在化疗后无１例阳性,化疗前后差异有统计学意义（Ｐ＝０.００１）。而对照组４５例外周血标本中均未检出ＰＩＰｍＲＮＡ的表达。ＰＩＰｍＲＮＡ的阳性表达与淋巴结转移、ＴＮＭ分期有关,而与患者年龄、肿瘤大小、ＥＲ、ＰＲ及Ｃ-ｅｒｂＢ-２等表达无关。结论：乳腺癌患者接受术后辅助化疗能够降低血液ＰＩＰｍＲＮＡ的阳性率,有望减少乳腺癌血液微转移的发生。     

t—PA,u—PA／uPAR及其抑制剂PAI—1在乳腺肿瘤的表达

目的　研究组织型纤溶酶原激活剂（ｔＰＡ） 、尿激酶型纤溶酶原激活剂（ｕＰＡ） 及其受体ｕＰＡＲ、１ 型纤溶酶原激活剂抑制剂ＰＡＩ１ 在乳腺肿瘤中的分布及酶学活性。　方法　用免疫组织化学ＡＢＣ方法,分析４ 种因子在乳腺肿瘤组织中的定位及半定量表达,用发色底物法测定乳腺肿瘤组织提取物中的ｔＰＡ 和ｕＰＡ 的活性。　结果　ｔＰＡ、ｕＰＡ、ＰＡＩ１ 主要分布在正常腺上皮及癌上皮细胞的胞质中,ｕＰＡＲ 则在癌细胞的胞膜及胞浆中;总ＰＡ 活性和ｔＰＡ 活性在乳腺良、恶性组织中均无显著性改变,但ｕＰＡ,ｕＰＡＲ 和ＰＡＩ１ 在乳腺癌组织中的表达明显高于良性组织及癌旁组织;ｕＰＡ／ｕＰＡＲ在转移灶中的表达较原发灶有不同程度的下降,而ＰＡＩ１ 在转移灶中却较原发灶中为高。　结论　ｔＰＡ 可能与乳腺癌的恶性表型无关。而ｕＰＡ／ｕＰＡＲ 与恶性表型相关;ｕＰＡ／ｕＰＡＲ 和ＰＡＩ１的相互制约及平衡在调节肿瘤原发灶癌细胞的移动中可能起到重要作用。     

阳离子脂质体——病毒增强质粒复合物在肝细胞癌基因治疗中的应用

目的建立一种简便有效的非病毒基因治疗方法。方法用构建的含人ＩＬ┐２基因的腺相关病毒（ＡＡＶ）增强质粒（ｐＡＩ┐ｈＩＬ┐２）与新型阳离子脂质体（Ｄｏｓｐｅｒ）以适当比例混合形成复合物（Ｄｏｓｐｅｒ┐ｐＡＩ┐ｈＩＬ┐２）。体外转染小鼠肝癌细胞系ＭＭ４５Ｔ．Ｌｉ┐２，评估了人ＩＬ┐２基因表达情况。应用直接瘤体内注射方法评估了此方法介导ＩＬ┐２基因的抗肿瘤作用。结果应用Ｄｏｓｐｅｒ┐ｐＡＩ┐ｈＩＬ┐２复合物体外转染ＭＭ４５Ｔ·Ｌｉ，其ＩＬ┐２表达水平每毫升可达１７５Ｕ／４８小时，最高表达时间在转染后第４天（２２５Ｕ／ｍｌ），第１２天仍可表达３０Ｕ／ｍｌ。瘤体内直接注射Ｄｏｓｐｅｒ┐ｐＡＩ┐ｈＩＬ┐２复合物后，瘤组织内可检出ＩＬ┐２ｍＲＮＡ，治疗组肿瘤生长明显减慢，小鼠生存率（转染后３０天）明显提高（Ｐ＜０．０１）。结论Ｄｏｓｐｅｒ┐ＡＡＶ增强质粒复合物能有效介导ＩＬ┐２基因表达，产生较好的抗瘤效应，此方法简便、更适合于临床应用     

乳腺肿瘤组织中Ha—ras基因蛋白表达与血清CA15—3水平的相关性及…

用免疫组化ＡＢＣ法和ＩＲＭＡ法分别检测了４２例乳腺癌和３０例良好性乳腺病患者组织中Ｈａ－ｒａｓ基因蛋白（ｒａｓｐ２１）的表达状况及血清ＣＡ１５－３，结果恶性病变组织中ｒａｓＰ２１呈过度表达，ｒａｓＰ２１的表达与细胞分化状态密切相关，不同淋巴吉转移状况的乳腺癌患者，其血清ＣＡ１５－３差异呈显著性，ＣＡ１５－３水平的变化与ｒａｓＰ２１表达状况显著相关，提示血清ＣＡ１５－３水平为监测乳腺癌病情变化的良性     

多药耐药相关蛋白在乳腺癌细胞中的表达

目的 观察多药耐药相关蛋白（ＭＲＰ）是否在临床乳腺癌组织中表达，了解ＭＲＰ在乳腺癌多药耐药现象中的临床相关性。方法 用ＲＴ－ＰＣＲ名免疫且化技术检测乳腺癌、乳腺纤维瘤组织中ＭＲＰ及ＭＲＰ ｍＲＮＡ的表达。结果 乳腺癌组织的ＭＲＰ阳性率的５６％，乳腺纤维瘤的２０％，阳性染色主要位于部细胞胞浆，胞膜次之。结论 ＲＴ－ＰＣＲ进一步表明术前化疗的乳腺癌组织ＭＲＰ ｍＲＮＡ表达明显强于乳腺纤维瘤及术前未受化     

人肿瘤裸鼠移植瘤株模型转移抑制基因nm23-H1mRNA的表达

目的ｎｍ２３基因与肿瘤移植瘤株转移的关系。方法应用地高辛标记的ｎｍ２３┐Ｈ１反义ｃＲＮＡ探针原位杂交方法对人克隆化肝癌，人骨肉瘤，肾癌三种不同肿瘤的裸鼠移植瘤株模型（Ｈ┐ＣＳ，Ｍ┐ＯＳ，Ｍ┐ＲＣＣ）中瘤内ｎｍ２３┐Ｈ１进行检测。结果ｎｍ２３Ｈ１ｍＲＮＡ在三模型中均有不同程度快的表达，其中有较强转移能力的Ｈ┐ＣＳ，和潜伏期短、侵袭生长速度快的Ｍ┐ＲＣＣ杂交信号５０％为弱阳性，而Ｍ┐ＯＳ仅有２５％为弱阳性。结论ｎｍ２３的弱阳性表达可能和Ｈ┐ＣＳ生长速度及转移倾向有一定联系。     

大肠癌组织中Ｎｅｕｒｏｐｉｌｉｎ-１的表达及其与肿瘤血管生成的相关性研究

目的：检测大肠癌组织中神经纤毛蛋白-１（Ｎｅｕｒｏｐｉｌｉｎ-１,ＮＲＰ-１）ｍＲＮＡ的表达情况及微血管密度（ＭＶＤ）值,探讨ＮＲＰ-１ｍＲＮＡ的表达与ＭＶＤ间的相关性,以期了解ＮＲＰ-１与肿瘤血管生成的关系。方法：取４５例大肠癌组织及相应正常大肠组织,采用半定量ＲＴ-ＰＣＲ方法检测其ＮＲＰ-１ｍＲＮＡ的表达情况,采用免疫组化染色法检测ＣＤ１０５标记的新生肿瘤血管,计数ＭＶＤ值。结果：４５例大肠癌组织中３９例可见ＮＲＰ-１ｍＲＮＡ的表达,阳性率为８７.０％;相应正常大肠组织仅３例表达,阳性率为６.７％。大肠癌组织中ＮＲＰ-１ｍＲＮＡ的表达程度与肿瘤的大小、浸润的深度、淋巴结转移呈正相关（Ｐ＜０.０５）,与肿瘤部位、组织学类型、分化程度、患者的性别及年龄无关（Ｐ＞０.０５）。大肠癌组织中ＮＲＰ-１ｍＲＮＡ的表达程度随ＭＶＤ值的增加而升高（ｒ＝０.３８,Ｐ＝０.０１）。结论：ＮＲＰ-１可能与大肠癌的肿瘤血管生成相关,ＮＲＰ-１ｍＲＮＡ高表达代表着大肠癌不良的生物学行为趋向。     

Comparative studies of TIMP-1 immunohistochemistry,TIMP-1 FISH analysis and plasma TIMP-1 in glioblastoma patients

Tissue inhibitor of metalloproteinases-1 (TIMP-1) has been associated with poor prognosis and resistance towards chemotherapy in several cancer forms. In a previous study we found an association between a low TIMP-1 tumor immunoreactivity and increased survival for glioblastoma patients, when compared to moderate and high TIMP-1 tumor immunoreactivity. The aim of the present study was to further evaluate TIMP-1 as a biomarker in gliomas by studying TIMP-1 gene copy numbers by fluorescence in situ hybridization (FISH) on 33 glioblastoma biopsies and by measuring levels of TIMP-1 in plasma obtained pre-operatively from 43 patients (31 gliomas including 21 glioblastomas) by enzyme-linked immunosorbent assay (ELISA). The results showed TIMP-1 gene copy numbers per cell ranging from 1 to 5 and the TIMP-1/CEN-X ratio ranging between 0.7 and 1.09, suggesting neither amplification nor loss of the TIMP-1 gene. The TIMP-1 protein levels measured in plasma were not significantly higher than TIMP-1 levels measured in healthy subjects. No correlation was identified between TIMP-1 tumor cell immunoreactivities and the TIMP-1 gene copy numbers or the plasma TIMP-1 levels. In conclusion, high immunohistochemical TIMP-1 protein levels in glioblastomas were not caused by TIMP-1 gene amplification and TIMP-1 in plasma was low and not directly related to tumor TIMP-1 immunoreactivity. The study suggests that TIMP-1 immunohistochemistry is the method of choice for future clinical studies evaluating TIMP-1 as a biomarker in glioblastomas.     

NSCLC患者血清TIMP-1和TIMP-2检测的临床价值分析

目的:探讨血清金属蛋白酶组织抑制剂-1(TIMP-1)和金属蛋白酶组织抑制剂-2(TlMP-2)水平在晚期非小细胞肺癌(NSCLC)诊断及疗效预后评价中的临床价值.方法:用酶联免疫吸附法(ELISA)检测26名健康人和20例良性肺病患者及82例晚期NSCLC患者化疗前后血清TIMP-1、TIMP-2水平,并作对比分析.结果:NSCLC组化疗前血清中TIMP-1、TIMP-2水平均明显高于健康组(P＜0.01)和良性肺病组(P＜0.05).NSCLC患者化疗有效组在化疗结束后7 d的血清TIMP-1、TIMP-2水平明显低于化疗前,P＜0.01;化疗无效组化疗前、后血清TIMP-1、TIMP-2水平差异无统计学意义,P＞0.05.单因素生存分析提示,血清TIMP-1水平与晚期NSCLC患者的预后有关,P＜0.05.结论:NSCLC患者血清TIMP-1水平是一个有价值的肿瘤标志和预后指标,检测化疗前后血清TIMP-1、TIMP-2水平对评估化疗疗效有一定的临床意义.     

TIMP-1 alters susceptibility to carcinogenesis

Tissue inhibitors of metalloproteinases (TIMPs) are a family of multifunctional proteins known to possess a broad range of biological activities, including inhibition of metalloproteinase activity, regulation of proliferation and apoptosis of a variety of cell types, and, depending on the context, differential regulation of angiogenic and inflammatory responses. Elevated mRNA expression of TIMP family members correlates with malignancy and clinical outcome in many human cancer types; however, a protective role for TIMPs also has been observed in various mouse models of human cancer. In the current study, we found distinct spatial-temporal expression patterns for the mRNA of TIMP family members in a mouse model of epithelial carcinogenesis [i.e., keratin 14-human papillomavirus 16 (K14-HPV16) transgenic mice]. To test the hypothesis that elevated expression of TIMP-1 functionally regulates epithelial carcinogenesis, we introduced a human TIMP-1 transgene into K14-HPV16 transgenic mice and assessed neoplastic progression. Results from these studies suggest that TIMP-1 enhances tumorgenicity by potentiating keratinocyte hyperproliferation and appearance of chromosomal aberrations in premalignant cells, thereby increasing their risk to undergo malignant conversion. In addition, TIMP-1 inhibits tissue gelatinolytic activity in tumor stroma, affects stabilization of collagen fibrils, but does not inhibit malignant conversion of dysplasias into carcinomas or development of metastases. The combined implications of these studies suggest that TIMP-1 is an important contributor to epithelial neoplastic progression and supports the concept that TIMP-1 exerts differential regulation on tissues in a stage-dependent manner.     

Clinical relevance of MMP-9, MMP-2, TIMP-1 and TIMP-2 in colorectal cancer

The main aim of this study was to evaluate the clinical relevance of Gelatinases in colorectal cancer (CRC). Ninety-five CRCs and their paired normal tissues were investigated to detect total levels of MMP-9, MMP-2, and the tissue inhibitors TIMP-1 and TIMP-2. Also, pro-MMP and MMP activity, and potential associations with clinical parameters were estimated. MMP-9, MMP-2 and TIMP-1 levels were greater in CRCs than in normal tissues, differences being significant for MMP-9 and TIMP-1. However, TIMP-2 showed significantly lower levels in tumour samples. Moreover, significant differences in the state of activation between gelatinases were found. TIMP-1 low levels were significantly associated with poor clinical outcome of patients. According to these data, different roles have to be attributed to MMP-2 and MMP-9 in CRC progression. Moreover, TIMP-1 level evaluation emerges as the main prognostic factor in relation to Gelatinases A and B activity in CRC.     

THE MANAGEMENT OF THE DISEASE

Occasionally, surgery is needed for particular complications in Stages I1 (B) and I11 GLWS. Splenectomy may become the only method for control of some cases of secondary hypersplenism (Grace and Mittieman, 1966). Intestinal obstruction, bowel perforation or haemorrhage or progressive spinal cord compression can demand an urgent operation. Resides surgical disorders, not related to Hodgkin's disease, will occur a t times.     

Adenovirus-mediated expression of TIMP-1 and TIMP-2 in bone inhibits osteolytic degradation by human prostate cancer

Matrix metalloproteinases (MMPs) are proteolytic enzymes that play critical roles in the pathogenesis of human cancers. Clinical trials using synthetic small molecule MMP inhibitors have been carried out but with little success. Tissue inhibitors of metalloproteinases (TIMPs) are endogenous inhibitors that block the extracellular matrix-degrading activity of MMPs. Here, we investigated the possibilities of genetically modifying human bones with TIMPs to create a high-TIMP bone microenvironment, which is hostile to metastatic prostate cancer cells using adenovirus-mediated gene transfer technology and SCID-hu end-organ colonization mouse model. Two strategies were used to achieve bone-specific TIMP expression: (i) ex vivo bone adenoviral infection followed by in vivo bone implantation; and (ii) ex vivo BMS cell infection followed by injection into in vivo implanted human fetal bones. PC-3 prostate cancer cells were injected into human fetal bones 4 weeks after implantation in SCID mice. In vitro, adenovirus-mediated expression of TIMP-1 or TIMP-2 in bone fragments inhibited MMP-2 activity, bone turnover and prostate cancer cell-induced proteolytic degradation as determined by gelatin zymography, calcium measurement and DQ protein quenched fluorescence assay, respectively. In vivo, immunohistochemistry confirmed TIMP-2 expression in AdTIMP-2-infected bone implants 4 weeks after implantation in SCID mice. Mice receiving AdTIMP-treated bone fragments showed significantly reduced PC-3-induced osteolysis, osteoclast recruitment and bone turnover in the implanted bones. We propose that adenoviral gene transfer of TIMP-1 and TIMP-2 can prevent the proteolytic activity of prostate cancer cells in bone and that enhancing anti-proteolytic defense mechanisms in target organs represents a promising form of prostate cancer gene therapy.     

食管前壁舌形切开预防食管胃吻合口狭窄的临床观察

２４６例食管胃吻合术随机分为甲，乙，丙三组，甲组吻合口后壁食管水平切开和胃间断缝合，前壁舌形切开，其舌形最高点距后壁２．０ｃｍ，然后与胃间断缝合，胃壁套埋吻合口的深度达２．５ｃｍ，前后壁亦与吻合口相平行，乙，丙两组则分别按传统的食管横切胃吻合和器史合，经半年以上随访，甲，乙，丙三组吻合口狭窄发生率依次为０％，４．８２％和１７．２２％。文中讨论了吻合口狭窄发生的因素及该手术方法预防吻合口狭窄的原理，