目的 了解E-钙粘蛋白在基因产物水平异质化表达对胃癌发生发展中的作用.方法 以35例新鲜胃癌标本和10例癌旁组织标本中提取的DNA为模板,采用PCR-单链构象多态性分析法对E-钙粘蛋白的exon8和exon9的基因突变进行筛选,对筛选阳性标本行DNA序列测定进行确证;采用免疫组化法检测以上标本的E-钙粘蛋白表达情况;采用SPSS 12.0统计软件进行数据分析.结果 35例胃癌标本中有3例检出E-钙粘蛋白exon8突变,突变阳性率为8.6％(3/35),exon9未发现基因突变,检测出的突变全部位于分化不良型胃癌标本中.6例分化型胃癌和10例癌旁组织中未检出基因突变.E-钙粘蛋白在癌与癌周黏膜、肿瘤分化程度及Lauren不同分型中的表达差异均具有统计学意义(P均＜ 0.05).结论 E-钙粘蛋白是参与胃癌发生、发展的重要分子,其表达与胃癌的分化程度、浸润性生长、组织结构完整性和破坏程度等生物学行为具有较显著相关性,其发生基因突变是导致胃癌发生、发展的重要因素.

目的 研究胃癌组织中线粒体DNA(mtDNA)D-loop区突变情况及其在肿瘤发生和发展中的作用.方法 选择17例胃癌及相应正常胃黏膜组织, 应用聚合酶链反应(PCR)对其mtDNA D-loop区进行扩增并测序, 将测序结果与线粒体文库中的Revised Cambridge Reference Sequence (rCRS)进行对比分析.结果 17例胃癌组织中共发现mtDNA D-loop区存在175个多态性变异,其中8个(4.6%)为新发现的变异.8例(47.1%)胃癌组织中共发现13次突变;突变热点集中在HV1(23.1%)和HV2(53.9%),并且HV2较HV1更易突变.mtDNA D-loop区突变率与胃癌分化程度、浸润深度、有无淋巴结转移、病人性别和年龄无关(P＞0.05).结论 mtDNA D-loop区尤其是其中的HV1及HV2是一个具有高度多态性和突变性的区域,在胃癌中突变率较高.

Objective To study the expressions of matrix metalloproteinase-10 (MMP-10), metalloproteinase 11 (MMP-11) and tissue inhibitor of metalloproteinase-2 (TIMP-2) in gastric carcinoma and explore the mechanism of invasion and metastasis of the cancer.MethodsExpressions of MMP-10, MMP-11 and TIMP-2 were studied via immunohistochemistry in specimens resected from 45 patients with gastric carcinoma and 20 paraneoplastic normal mucosa (PNM).ResultsDifferences in expressions of MMP-10, MMP-11 and TIMP-2 were significant between cancer mucosa and PNM (χ2=4.30-6.37,P0.05). Expressions of MMP-10 and MMP-11 were both correlated with tumor size, depth of tumor invasion, LN metastasis and TNM stage (χ2=6.46-15.91;P0.05,0.01); expression of TIMP-2 was correlated with LN metastasis (χ2=4.91,P0.05), and that of MMP-10 and MMP-11 were positively correlated with that of TIMP-2 (γ=0.29,0.34;P0.05).ConclusionThe abnormal expressions of MMP-10, MMP-11 and TIMP-2 are closely related with the metastasis and that can be used as important parameters for the judgement of biological behaviour and prognosis of gastric carcinoma.

Objective To explore the relationship between the expressions of matrix metalloproteinase 3, 7 (MMP3, MMP7) and tissue inhibitor of metalloproteinase 1 (TIMP1) and the biological behavior of gastrointestinal stromal tumor (GIST), in order to provide the theoretical basis and probable indicators for predicting the metastasis potentiality of the tumor. MethodsThe expressions of MMP3, MMP7 and TIMP1 in 68 GIST samples were studied by immunohistochemistry and their association with histo-morphology and biological behavior analyzed statistically.ResultsThe positive expressions of MMP3, MMP7 and TIMP1 increased with the increase of the Fletcher degree (r=0.299-0.354,P<0.01). The differences of the positive expressions among the three items were significant when the tumor size was ≤5 cm and >5 cm in diameter (χ2=4.73-5.82,P<0.05). The positive expressions of MMP3 and TIMP1 were different with or without metastasis (χ2=4.57,4.61;P<0.05). The differences between the expression degree of MMP3 in GIST with expansive growth and infiltrating growth, and of MMP3 in GIST with pathological mitotic figures counted ≤10/50HPF and >10/50HPF were also significant (χ2=4.69,5.81;P<0.05). ConclusionMMP3, MMP7 and TIMP1 are closely related with the ability of invasion and metastasis of GIST, and the imbalance among the three of them is a crucial factor to progress of malignancy of GIST. Detection of these three parameters can be regarded as a level of significance in assessment of the biological behavior of malignancy.

Objective To explore the relationship between DNA ploidy together with p53 expression and the biologic behaviors of peripheral nerve sheath tumors and study their clinical significance in differential diagnosis.MethodsFlow cytometry was used to analyze DNA ploidy in paraffin embedded tissue samples from 65 cases of benign and malignant peripheral nerve sheath tumors (MPNST). P53 protein expression was examined with immunohistochemistry.ResultsAll of the 23 cases of benign tumors were diploid, while four out of 42 MPNST patients were aneuploid. The difference was insignificant between the benign and MPNST. But the values of SPF and PI between the benign and MPNST groups were significantly different (χ2=28.674,23.730;P<0.001); Furthermore, the SPF and PI values were positively correlated with the histological grade of MPNST (rs=0.418;0.347;P<0.01,0.05). P53 protein expression in MPNST was significantly higher than that in the benign peripheral nerve sheath tumors (χ2=34.719,P<0.001). The intensity of P53 expression was positively correlated with the malignancy of MPNST (rs=0.326,P<0.05), as well as with the positive rate of aneuploidy, the values of SPF and PI (rs=0.318-0.354,P<0.05). ConclusionThe SPF and PI values were gradually increased during the transformation of benign peripheral nerve sheath tumors into MPNST. Therefore, SPF and PI may be used as indicators for differential diagnosis of benign and MPNST. P53 expression is a valuable factor for differential diagnosis between benign and MPNST, and for estimating the malignancy of MPNST.

Objective To investigate the expression of p53,p27 and mdm2 protein in gastrointestinal stromal tumor(GIST),and discuss the relationship between their expression and clincopathological characteristics.Methods Immunohistochemical staining was used to detect the expression of p53,p27 and mdm2 in 68 cases of GIST.Their relations with histo-morphology and biological behavior were analyzed statistically.Results In 68 cases GIST,the positivity of p53,p27 and mdm2 protein were 75%,54.41% and 27.94% respectively.The expression of p53 protein was inversely correlated with the fletcher degree,morphologic variant(P0.05) and tumor diameter(P0.01) of GIST.The expression of p27 protein was inversely correlated with the fletcher degree(P0.05) and tumor diameter of GIST(P0.01).The expression of mdm2 protein was inversely correlated with the morphologic variant(P0.01) of GIST.Conclusion These data shows the expression of p53,p27 and mdm2 are related to the ontogenesis and progression of GIST.p53,p27 and mdm2 may be prognostic indicators in GIST.

【Objective】To investigate the expression of Elf-1,Survivin and their clinical significance in non-small-cell lung cancer(NSCLC).【Methods】The expression of Elf-1,Survivin was detected immunohistochemistri-cally by PV9000 method with tissue chip contained 72 cases of NSCLC and 10 cases normal lung tissue.【Results】 Elf-1 and Survivin were both expressed heterogeneously in NSCLC.The positive rate of Elf-1 was 73.61% in NSCLC group and 2/10 in normal lung tissue,and it was obviously different between the two groups(P <0.01).The expressive intensity of Elf-1 was related to histopathological grading(χ2 =8.274,P =0.013),lymph node metastasis(Z=-3.316,P =0.001) and clinical staging(χ2 =24.127,P =0.000).The positive rate of Survivin was 61.11% in NSCLC group and 1/10 in normal lung tissue,and it was also obviously different between the two groups(P <0.01).The expressive intensity of Surviving was related to histopathological grading(χ2 =8.513,P =0.012),lymph node metastasis(Z =-5.125,P =0.000)and clinical staging(χ2 =20.570,P =0.000).Positive correlation was found between the expressive intensity of Elf-1 and Survivin(rs=0.255,P =0.012).【Conclusions】The expression of Elf-1 and Survivin may be correlate with the occurrence and progression of NSCLC,and exert synergistic effect in this progress.United detecting the expression of Elf-1 and Survivin may be served as the molecular biological index of evaluating invasion and metastasis,and may be usefull for early diagnosis of NSCLC.

OBJECTIVE:To study the expression of heparanase(HPSE)in pancreastic cancer and the correlation between HPSE and angiogenesis and prognosis.METHODS:Immunohistochemistry was used to detect heapranase and microvessel density(MVD)in 47 cases of pancreatic cancer,24 adjacent tissues of cancer and 10 normal tissues,and the correlation of the expression of HPSE protein with the clinicopathological charac teristics were investigated.In addition,CD34 was detected and the association of the HPSE expression with microvessel density(MVD)and extent of proliferation of pancreatic carcinomas was analysised.RESULTS:The rate of the expression of HPSE protein in 47 cases of pancreatic cancer tissues(59.6%,28/47)was significantly higher than those in the adjacent tissues of cancer(20.8%,5/24)and normal tissues(20.0%,2/10),χ2=9.59,3.90;P=0.002 0,0.0487 1.It was shown that HPSE protein expression did not significantly correlate with the size and histological grade of pancreatic cancer,χ2=1.09,2.89,P=0.580 4,0.235 8,but it correlated with the invasion,metastasis and survival time of the patients,χ2=6.52,9.25,6.19,P=0.010 7,0.002 4,0.012 8.The expression of HPSE protein was positively associated with MVD of pancreatic carcinomas,t=5.09,P=0.000 0.CONCLUSIONS:The expression of HPSE protein may play an important role in the growth,invasion,metastasis and angiogenesis of pancreatic cancer.Pancreatic cancer with a high proliferating activity tends to express HPSE.HPSE may be a valuable marker to evaluate biological behavior and prognosis of pancreastic cancer.

Objective:Through studying the expressions of PTEN in malignant peripheral nerve sheath tumors (MPNST) and analyzing the DNA ploidy by the flow cytometry from the level of single cell, to explor the relations between them and the biologic behaviors of MPNST.Methods:Using the method of immunohistochemistry, 65 cases of benign or malignant peripheral nerve sheath tumors were selected to examine PTEN indicator, all the samples were paraffin embedded. At the same time, the DNA ploidy of all these samples by flow cytometry were analyzed. Results:PTEN was expressed differently between benign and malignant peripheral nerve tumors and the difference was significant (χ2=17.628,P0.001). The stronger expression of PTEN was positively related to the degree of the malignancy of MPNST (rs=0.067,P=0.699). There were 4 aneuploidy in 42 cases of MPNST, and all the aneuploidy was low aneuploidy(DI0.90); SPF and PI index were also different between the benign and malignant peripheral nerve tumors, the differences were significant(χ2=28.674,P0.001;χ2=23.730,P0.001); SPF and PI index were also positive correlations with the histological grade of MPNST(rs=0.418,P0.01;rs=0.347,P0.05); The degree and intension of the expression of PTEN was positively correlated to the three indexes of DNA ploidy analysis.Conclusion:PTEN can be used as a worthy factor to estimate the property of benign and malignant peripheral tumors and as a valuable indicator to estimate the degree of malignancy of MPNST. The appearance of the aneuploidy and high SPF,PI index can predict the malignant degree(progression and heterogeneity) of MPNST.

Objective To evaluate the diagnosis value of F-PSA/T-PSA on early for prostate cancer(PCa) with prostate specific antigens(PSA) 4.0 μg/L.Methods The free and total PSA in 136 normal male volunteers(normal control),87 PCa patients with T-PSA≥4.0 μg/L and 12 patieats with T-PSA4.0 μg/L were measured by electrochemiluminescence immunoassay.The ratios of F-PSA/T-PSA were therefore obtained.Results The T-PSA, F-PSA and F-PSA/T-PSA consisted of 1.43±1.13 μg/L,0.44±0.34 μg/L,0.36±0.15 in cases of normal control group;31.25±20.68 μg/L,4.21±3.12 μg/L,0.17±0.08 in 87 cases with T-PSA≥4.0 μg/L PCa cases;2.56±1.11 μg/L,0.43±0.29 μg/L,0.18±0.09 in 12cases with T-PSA4.0 μg/L PCa cases.There is significant difference of T-PSA,F-PSA/T-PSA was observed between the normal control group and 12 PCa cases with 4.0 μg/L(P0.01).Significant difference of T-PSA、 F-PSA and no difference of F-PSA/T-PSA between Pca with T-PSA≥4.0 μg/L and Pca with T-PSA 4.0 μg/L.The cut off of F-PSA/T-PSA were ranked as 0.10,0.11～0.15,0.16～0.20,0.21～0.25,0.26～0.30,0.31.The 75% and 78.16% prevalence of PCa were observed in patients with T-PSA4.0 μg/L PCa,T-PSA≥4.0 μg/L PCa respectively when F-PSA/T-PSA cutoff was less than 0.20(P0.05).Conclusion The analysis of F-PSA/T-PSA is valuable for early diagnosis prostate cancer for patients with serum T-PSA≥4.0 μg/L and as low as 4.0 μg/L.

Objective To observe the cytological heterogenesis of glioma and analyze its effect on the tumor therapy and prognosis.Methods The data of 150 patients with glioma resected in this hospital,from January 2005 to June 2006,were collected and analyzed histologically.Results Sixty-nine gliomas(46%) showed more than two kinds of cellular differentiation.Astrocytoma(5%),anaplastic astrocytoma(77%),oligodendroglial tumor(26%),glioblastoma(60%) and all the seven-mixed gliocytomas had more than two kinds of cellular differentiation and this type of differentiation was also seen in six of eight recurrent astrocytomas.Conclusion Cellular heterogenesis of gliomas shows close relationship with the progression of the tumors.A further study of the tumor differentiation may bring about an active effect on the therapy and prognosis of the disease.

由国际癌症研究机构(IARC)最新出版的2004年版WHO肿瘤分类<泌尿系统和男性生殖器官病理和人体遗传学>一书中,MATIGNONI和AMIN将肾血管平滑肌脂肪瘤(AML)的组织学分型、定义、病因学、遗传学、良恶性评估和预后进行了较1998年版更为详细的描述.

Objective To investigate the expression of CD34 of microvessel density(MVD) and the relationship among MVD,clinicopathological features and cell proliferation in patients with endometrial adenocarcinoma. Methods The expressions of CD34 and Ki-67 in 20 cases of normal endometrium and 70 cases of endometrial adenocarcinoma were detected by immunohistochemical EnviSion method. Results MVD value in the endometrial adenocarcinoma was significantly higher than that in the normal endometrium(t=12.67,P<0.001).There was no relationship between the value of MVD and patient's age,pathological grade(t=1.673,F=2.09,P>0.05),but it was related to clinical stage,myometrial invasion and lymph node metastasis(F=5.77;t=5.650,6.445;P<0.01,0.001).Expression of Ki-67 was positively correlated with MVD(t=2.522,P<0.05). Conclusion MVD is associated with the growth,invasion and metastasis of endometrial adenocarcinoma.Combination detection of MVD and Ki-67 can be used as a parameter to judge the biological behavior and prognosis of endometrial adenocarcinoma.

Objective To study DNA ploidy and expression of MMP-9 and TIMP-2 in patients with gastric carcinoma and explore the mechanism of invasion and metastasis of the disease. Methods Immunohistochemistry was used to detect the expressions of MMP-9 and TIMP-2 in specimens from 156 patients, including specimens of the carcinoma (99), of surrounding mucosa (16), of distant metastases (16) and of metastastic lymph node (25). Flow cytometry DNA ploidy and S-phase fraction (SPF) analysis was done in specimens from 57 patients, including 47 specimens of the cancer, of adjacent mucosa (6) and of distant metastatic focus (4). Results Expression of MMP-9 was correlated with LAUREN classification, BORRMANN classification, LN metastasis and TNM stage (χ2=6.65-8.03;P0.05,0.01). Expression of TIMP-2 was significantly correlated with BORRMANN classification and LN metastasis (χ2=5.01-7.05,P0.05). Positive correlation between MMP-9 and TIMP-2 was observed (r=0.22,P0.05). The differences of expression of MMP-9, the rate of DNA heteroploid and SPF were significant between cancer mucosa and non-cancer mucosa. Conclusion The abnormal expressions of MMP-9 and TIMP-2, DNA heteroploid and higher SPF are involved in the progression and heterogeneity of gastric carcinoma.

目的 研究Wnt信号中β-catenin基因第3外显子突变及蛋白异常表达在胰腺实性-假乳头状瘤(SPTs)发生中的作用.方法①对15例SPTs石蜡包埋组织进行DNA提取,PCR扩增,PCR产物纯化、测序,对β-catenin基因第3外显子进行突变分析.②采用免疫组化PV6000法检测SPTs组织中β-catenin蛋白的表达.结果①β-catenin基因第3外显子在15例SPTs组织中有12例发生了一个碱基的错义突变,突变率为80.0%,表现在密码子32位(5例)、33位(3例)和37位(4例).②15例SPTs组织中有13例β-catenin蛋白呈细胞核表达,异常表达率为86.7%.结论Wnt信号中β-catenin基因突变及蛋白核内聚集在SPTs发生中起重要作用。

患者女,28岁.因头晕心慌在心内科就诊时发现腹部肿块,经B超检查发现肾脏巨大肿瘤伴瘤内出血,于2005年1月16日入院.体检:患者自6岁开始有多次癫痫发作,青春期后发作逐渐减少,无家族史.

ObjectiveTo investigate the expressions of P-gp, TOPOⅡ,GST-π and LRP in infiltrating ductal carcinoma of breast and evaluate their clinical significance.MethodsP-gp,TOPOⅡ,GST-π and LRP were measured in the patients (n=80) by immunohistochemistry using monoclonal antibodies against these proteins.ResultsThe positive rate of P-gp,TOPOⅡ,GST-π and LRP was 40%,55%,65% and 80%,respectively.The interrelationship among their expressions was not found. ConclusionA combined analysis of the expression of P-gp,TOPOⅡ,GST-π and LRP in breast cancer may contribute to the drug selection for chemotherapy and prediction of the prognosis of the disease.

Objective To investigate the relationship between Preoperative epilepsy and the biological behaviour of astrocytoma, and the relationship between Preoperative epilepsy and the prognosis of patients. Methods Data of 47 patients who were treated at the neurosurgery of the affiliated hospital of Qingdao university between 1995 and 2000 were collected and analyzed . Results The incidence of Preoperative epilepsy between astrocytomas of different biological behaviour are significantly different(P<0.01), and the survival between the patients with preoperative epilepsy and the patients without preoperative epilepsy are significantly different(P<0.01). Conclusion The incidence of Preoperative epilepsy is high in lower invasive astrocytoma, which can be helpful to estimate the survival.

Objective To investigate the expression of Ki-67 protein and caspase-3 in endometrial adenocarcinoma and its relationship with the parameter of clinical pathology and to interpret their role in the genesis and development of endometrial adenocarcinoma. Methods The expression of Ki-67 protein in endometrial adenocarcinoma and caspase-3 was detected by immunohistochemical EnviSion method in 20 cases of normal endometrium and 70 of endometrial adenocarcinoma. Results The positive expression rate of Ki-67 protein in endometrial adenocarcinoma was significantly higher than that in normal endometrium(Hc=27.81,P0.01);the positive expression of caspase-3 was markedly lower in endometrial adenocarcinoma than that in normal endometrium(Hc=27.81,P0.01).There was no relationship between Ki-67 protein expression and patient's age and clinical stage(Hc=3.590,5.888;P0.05),but it was related to pathological grade,myometrial invasion and lymph node metastasis (Hc=8.504,4.778;uc=2.963;P0.05,0.01).The expression of caspase-3 did not correlate with patient's age,clinical stage,myometrial invasion,and lymph node metastasis,but it correlated with pathological grade(Hc=7.320,P0.05).Expression of Ki-67 was correlated negatively with caspase-3(rs=-0.241,P0.01). Conclusion The expression of Ki-67 is closely related to the infiltration and metastasis of endometrial adenocarcinoma,acting as a negative feedback regulation,Ki-67 and caspase-3 may be involved in common in the genesis and development of endometrial adenocarcinoma,and in the pathological process of cell apoptosis and proliferation.

Objective To investigate the expression of β-catenin and MMP7 in various grade of glioma and the clinical significance.Methods Tissue microarray were constructed including 89 glioma and 6 normal brain tissues.Two step immunohistochemical staining was used to examine the expression of β-catenin and MMP7 in various grade of glioma and normal brain tissues.Results Normal brain tissues showed negative expression forβ-catenin and MMP7.A higher expression of β-catenin and MMP7 could be observed with the increasing grade of glioma.The expression of β-catenin and MMP7 were significantly different in low-grade and high-grade glioma(P0.01,P0.05 respectively).The expression of β-catenin had close correlation to that of MMP7 in glioma (P0.01).Conclusion The accumulation of β-catenin in cytoplasm and nuclear could promote the expression of MMP7,which might be associated with tumorigenesis,development and aggression of glioma.The expression of β-catenin suggested the malignant degree and grade of glioma.