良性前列腺增生的形态学改变与ARmRNA表达差异

采用直接法原位ＰＣＲ技术对４例ＮＡＰ和１０例ＢＰＨ组织中ＡＲｍＲＮＡ表达作了检测分析，并对其在ＮＡＰ与ＢＰＨ的表达作了定量比较分析。结果表明∶（１）ＮＡＰ及ＢＰＨ标本ＡＲｍＲＮＡ表达均显示阳性。阳性信号主要定位于靠近核膜的胞浆当中，主要分布在前列腺上皮细胞、间质细胞。（２）前列腺上皮组织中ＡＲｍＲＮＡ表达具区域异质性的特点；部分增生前列腺的腺上皮基底层（ＢＣＬ）细胞阳性表达增强，ＮＡＰ组未发现此现象。（３）图像分析表明无论是ＮＡＰ还是ＢＰＨ，ＡＲｍＲＮＡ在腺上皮的阳性表达强度均强于间质。而ＮＡＰ与ＢＰＨ之间阳性表达强度无显著差异。     

凋亡相关基因bcl-2、bax蛋白在前列腺良、恶性病变中的表达及意义

探讨细胞凋亡相关基因 bcl- 2、bax在前列腺良、恶性病变中的表达及意义。方法　应用免疫组织化学 SP法检测 2 9例前列腺癌 ( Pca)组织 ,35例前列腺增生 ( BPH)组织和 1 7例正常前列腺 ( NP)组织中 bcl- 2、bax蛋白表达。结果　 Pca组和 BPH组 bcl- 2蛋白表达阳性率明显高于 NP组 (均 P <0 .0 5) ,而 Pca组与 BPH组阳性率无显著性差异。 bax蛋白在 Pca、BPH、NP各组表达阳性率无显著性差异。Pca组织中 bcl- 2蛋白表达与病理分级、血 PSA浓度呈正相关 (均 P <0 .0 5) ,而与临床分期无显著相关性 ;bax蛋白表达与 Pca临床、病理指标相关分析均无显著意义。在 Pca、BPH组织中 bcl- 2和 bax蛋白两者表达呈负相关。结论　由于 bcl- 2蛋白过量表达引起的 bcl- 2 / bax失平衡 ,在 BPH、Pca发生、发展过程中起着重要作用 ,bcl- 2蛋白检测可以作为判断 Pca生物学特性的重要指标。     

THE STUDY ON RELATION BETWEEN CD1a^＋ CELLS AND INFILTRATIVE CD3^＋ AND CD8^＋ CELLS IN RENAL TISSUE OF GLOMERULONEPHRITIS

Objective To discuss the role of dendritic cells (DCs) in cellular immunity pathogenesis of glomerulonephritls (GN). Methods 114 patients with GN were selected randomly and divided into two groups,primary GN (pGN) and secondry GN (sGN). CDIa^ , CD3^ and CD8^ cells in bioptic renal tissues were examine dy immunohistochemically. The distribution of CDla cells and the infiltration of CD3^ and CD8^ cells in renal tissues were observed. Results There was no significant difference of CDIa^ , CD3^ , and CD8^ cells between pGN and sGN group (P＞0. 05). CDIa^ cells had significant positive correlation with the infiltrative CD3^ and CD8 cells,respectively (P＜0.01). The infiltrative CD3^ cells had significant positive correlation with the CD8 cells in the same area, respectively (P＜0.01). CD1a^ cells, CD3^ cells infiltrating in both glomeruli and renal interstitial tissues, and CD8^ cells only infiltrating in renal interstitial tissues, all of them had significant positive correlation with the degree of glomerular proliferation, respectively (P＜0.05). The infiltrative CD3^ and CD8^ cells in renal interstitial tissues had significant positive correlation with the degree of glomerular sclerosis and the lesion degree of renal tubule and interstitial, respectively (P~ 0. 05). There were significant positive correlation between CDla cells and the lesion degree of renal interstitial (P＜0.05). Collusion DCs could activate T lymphocyte by presenting antigen, then the activated T lymphocyte participate in the pathogenesls of GN through releasing cytokine and/or directly damaging the renal tubule and interstitial, which produce more serious glomeralar lesion.     

EXPRESSION OF ENDOGENOUS OUABAIN IN MULTIPLE ADRENAL TUMORS

Ａｄｒｅｎａｌｓ ,ｗｈｉｃｈｃａｎｋｅｅｐｔｈｅｂａｌａｎｃｅｏｆｉｎｎｅｒｅｎｖｉｒｏｎｍｅｎｔ,ａｒｅｉｍｐｏｒｔａｎｔｅｎｄｏｃｒｉｎｅｓ ,ａｎｄｌｏｔｓｏｆｄｉｓｅａｓｅｓａｒｅｒｅｌａｔｅｄｔｏｔｈｅｉｒｆｕｎｃｔｉｏｎ .ＴｈｅｐｒｅｓｅｎｃｅｏｆａｎｅｎｄｏｇｅｎｏｕｓＮａ ,Ｋ - ｐｕｍｐｉｎｈｉｂｉｔｏｒｉｎｍａｍ ｍａｌｓ ,ｗｈｉｃｈｃａｎａｆｆｅｃｔｒｅｎａｌＮａ ｅｘｃｒｅｔｉｏｎ ,ｖａｓｃｕｌａｒｃｏｎｔｒａｃｔｉｌｉｔｙ ,ａｎｄｎｅｒｖｏｕｓｒｅａｃｔｉｖｉｔｙ ,ｈａｓｂ…     

THE CLINICAL SIGNIFICANCE OF PERCENT FREE PSA IN DIFFERENTIATING PROSTATE CANCER AND BENIGN PROSTATE HYPERPLASIA

rostatecancer (Pca)isoneofthecommonestmalignanttumorsofmaleinEuropeandAmerica .ThemorbidityofPcainourcountryismuchlowerthaninothercountries,butrecentlyitwasincreasedtotwofold[1] .Althoughtheprostatespecificantigen(PSA)isthemostusefultumormarkeravailableforthediagnosisofPca ,nevertheless ,lackssufficientsensitivityandspecificity .Peoplenow putforwardtheconceptofpercentfreePSA (fPSA)andhasbe comethenewdirectionofinvestigation .However ,therestillhavesomeproblemunclearatpresent,es peciallyt…     

bcl-2基因蛋白在外阴营养不良及外阴癌的表达

利用细胞凋亡抑制基因ｂｃｌ－２单克隆抗体及免疫组化ＬＳＡＢ法,对正常外阴皮肤、慢性外阴营养不良、不典型增生和外阴癌组织进行标记,发现ｂｃｌ－２蛋白在正常外阴皮肤基底层中有弱阳性表达;在增生型营养不良、不典型增生及外阴癌组织中表达率有不同程度增高,与外阴硬化苔癣型和混合型营养不良有显著差异（Ｐ＜０．０５,Ｐ＜０．０５,Ｐ＜０．０１）;而增生型与不典型增生ｂｃｌ－２基因蛋白表达无明显差异（Ｐ＞０．０５）;外阴癌的表达率最高;该基因蛋白表达与外阴癌细胞分化程度有关,与临床分期无关。     

Tenascin、CD34在胆管癌组织中的表达及临床意义

目的探讨Tenascin、CD34在胆管癌的发生、发展中的作用和意义。方法收集我院1995年-2003年期间切除的胆管癌标本33例和正常胆管组织4例,采用免疫组织化学S-P方法,检测其Tenascin、CD34的表达情况。结果Tenascin在胆管癌组织中的阳性表达率为81.8%(27/33),其中病理分级Ⅰ级、Ⅱ级、Ⅲ级的阳性率分别为71.4%、82.4%和88.9%,但无显著性统计学差异(P>0.05);在正常胆管组织中未检测到Tenascin表达。CD34在胆管癌组织中广泛表达,在正常胆管组织中仅有少量表达,两者相比具有显著统计学差异(P<0.05)。结论Tena-scin可以作为胆管癌的肿瘤标志应用于临床诊断,CD34可作为预测肿瘤转移的一项重要指标。     

FPSA/TPSA比值在前列腺癌诊断中的意义

目的　评价FPSA/TPSA比值在前列腺癌诊断中的作用。方法　 32例前列腺癌 (PC)及97例前列腺良性增生 (BPH)患者 ,经活检穿刺或 (和 )手术病理确诊 ,采用酶联免疫方法测定血清中总前列腺特异性抗原 (TPSA) ,游离前列腺特异性抗原 (FPSA)浓度 ,计算FPSA/TPSA(F/T)比值。结果　TPSA浓度在 4.0～ 1 0 .0 μg·L- 1 范围时 ,其浓度两组间无显著性差异 (P >0 .0 5 ) ,而F/T比值则有显著性差异 (P <0 .0 1 ) ;测得F/T比值在 0 .2 5水平时 ,总有效率最大( 62 .9% )。结论　TPSA单独作为前列腺癌和前列腺良性增生鉴别诊断的指标存在局限性。当TPSA水平在 4.0～ 1 0 .0 μg·L- 1 范围内 ,应用F/T比值较PSA为优。F/T比值在 0 .2 5界值时诊断PC和BPH最有效     

DETECTION AND SIGNIFICANCE OF HBV IN RENAL TISSUE OF HBV ASSOCIATED GLOMERULONEPHRITIS PATIENTS

ＳｉｎｃｅＣｏｍｂｅｓ[1] ｆｉｒｓｔｌｙｒｅｐｏｒｔｅｄｏｎｅｃａｓｅｏｆｔｈｅＨＢＶ ａｓｓｏｃｉａｔｅｄｍｅｍｂｒａｎｏｕｓｎｅｐｈｒｏｐａｔｈｙ (ＨＢＶ ｍＧＮ )ｉｎ 1971,ｔｈｅｒｏｌｅｏｆｈｅｐａｔｉｔｉｓＢｖｉｒｕｓ (ＨＢＶ)ｐｌａｙｅｄｉｎ ｇｌｏｍｅｒｕｌｏｎｅｐｈｒｉｔｉｓｗａｓ ｇｅｎｅｒａｌｌｙ ｐａｉｄｃｌｏｓｅａｔｔｅｎｔｉｏｎｗｈｅｎｉｔｓｍｅｃｈａｎｉｓｍｗａｓｉｎｖｅｓｔｉｇａｔ ｅｄ .Ｉｔｒｅｍａｉｎｓｕｎｃｅｒｔａｉｎａｔｐｒｅｓｅｎｔ,ｈｏｗｅｖｅｒ ,ｈｏｗＨＢＶｐｌ…     

狼疮性肾炎肾活检组织中PCNA与Bcl-2的表达及意义

目的　观察狼疮性肾炎 (lupusnephritis ,LN)肾组织内凋亡调控基因Bcl 2及增殖细胞核抗原 (PCNA)的表达 ,探讨凋亡在LN发病中的作用。方法　应用ELPS(enhancedlabelled polymersystem)检测 2 9例LN肾穿刺活检组织PCNA和Bcl 2的表达 ,并与正常组进行对比分析。结果　LN组肾小球内PCNA阳性细胞数与正常对照组肾组织比较 ,有显著性差异 (P <0 .0 0 1)。LN组肾小球内Bcl 2阳性表达细胞明显高于正常对照组 (P <0 .0 0 1) ,阳性表达细胞见于系膜细胞、毛细血管内皮细胞和细胞性新月体内。LN组肾小球Bcl 2阳性细胞数与PCNA阳性细胞数、肾小球细胞总数有显著正性线性相关 (r =0 .70 1.P <0 .0 1;r =0 .5 5 0 ,P <0 .0 1)。结论　LN肾组织肾小球内细胞高增殖和Bcl 2高表达 ,提示细胞增殖与凋亡调控异常均参与了LN肾小球的增生性病变     

中性粒细胞硷性磷酸酶(NAP)测定在白血病鉴别诊断中的价值

本文报导了101例白血病及26例感染患者的髓细胞NAP活性测定。结果急非淋、急淋与淋巴瘤细胞白血病间无显著性差异,均可呈强阳性反应,亦可为阴性,慢粒髓细胞,NAP活性明显减低,慢粒急变者则明显升高,其酶活性与感染及类白血病反应相似。故NAP活性测定呈强阳性反应者,应首先考虑类白血病反应,但不能完全排除白血病之可能,特别是慢粒急变的初发型及CNL,应结合临床进一步探讨其它鉴别诊断之依据。NAP活性测定不能作为鉴别急淋与急非淋的可靠依据。     

EXPRESSION OF MATRIX METALLOPROTEINASE-9 IN HUMAN ABDOMINAL AORTIC ANEURYSMAL TISSUES

Objective To study the effects of MMP-9 （Matrix Metalloproteinase-9, MMP-9） in the pathogenesis of abdominal aortic aneurysms （AAAs） by localizing the expression of MMP-9 in the aneurysmal tissues. Methods By , the frozen sections （5μm） with aneurysmal tissues （n = 10） were incubated with MMP-9 antibody-added agents, then the sections were stained and observed under the microscope to localize the expression of MMP-9, which displayed a brown precipitate within the arterial walls. The normal arterial wall tissues（n= 10） and the diseased arterial wall tissues from the arterial occlusive diseases （AODs） （n= 15） were also immunized exactly the same way as control. Results A quantity of positive granules which appeared within the aortic media showed the strong expression of MMP-9 in the AAAs, with the positive rate reaching 95 % （19/20）, while no expression of MMP- 9 was observed in the normal artery. However, the scattered distributed positive granules were seen within the arterial wall of some cases of the AOI）s, implying the weak positive expression of MMP-9 in this disease with the positive rate of 26.7% （4/15）. There was a significant difference of the expression of MMP-9 within the arterial wall between the AAAs and AODs（P〈0.01 ）. Conclusion High expression of MMP-9 within the aortic media faciliatates the degradation of collagen and elustin fibres and subsequent dilation of the aortic artery , thus playing an important role in the pathogenesis of AAAs. To refrain MMP-9 from enhanced expressing within the aortic wall is of clinical significance in the prevention and treatment of AAAs.     

STUDY ON B7-1 PROTEIN‘S EXPRESSION IN BIOPSIES IN RENAL TRANSPLANTATION

Ａｌｌｏｇｒａｆｔｒｅｊｅｃｔｉｏｎｉｓａｎｉｍｐｏｒｔａｎｔｆａｃｔｏｒｏｆｉｎ ｆｌｕｅｎｃｉｎｇｔｈｅｌｏｎｇｔｅｒｍｓｕｒｖｉｖａｌｏｆａｌｌｏｇｒａｆｔｉｎｈｕ ｍａｎｏｒｇａｎｔｒａｎｓｐｌａｎｔａｔｉｏｎ .Ｉｍｍｕｎｏｌｏｇｉｃａｌｔｏｌｅｒａｎｃｅｉｎｄｕｃｔｉｏｎｉｓａｍａｉｎ ｐａｒｔｉｎｔｈｅｒｅｓｅａｒｃｈｏｆｏｒｇａｎｔｒａｎｓｐｌａｎｔａｔｉｏｎ .Ｔｏｓｔｕｄｙｔｈｅｉｍｍｕｎｏｌｏｇｉｃａｌａｎｄｍｏｌｅｃｕｌａｒｍｅｃｈａｎｉｓｍｏｆｒｅｊｅｃｔｉｏｎａｎｄｓｅｅｋ…     

Bcl-2和Bax蛋白表达与非小细胞肺癌临床病理特征的关系

目的　探讨 Bcl- 2和 Bax蛋白表达与非小细胞肺癌 (NSCLC)临床病理特征的关系。方法　应用免疫组化 SP法检测了 30例正常肺组织和 61例 NSCLC中 Bcl- 2和 Bax蛋白的表达。结果　 NSCLC组织中 Bcl- 2蛋白表达率 (57.38% )显著高于正常肺组织 (1 3.33% ) (P <0 .0 1 ) ,而NSCLC组织中 Bax蛋白表达率 (45.90 % )显著低于正常肺组织 (80 .0 0 % ) (P <0 .0 1 ) ,鳞癌和腺癌中 Bcl- 2和 Bax蛋白表达率均无显著差异 (P >0 .0 5) ,Bcl- 2蛋白表达率与 NSCLC病理分级及淋巴转移潜能呈正相关 (P <0 .0 5) ,与 P- TNM分期无关 (P >0 .0 5) ,Bax蛋白表达率与NSCLC病理分级、P- TNM分期及淋巴转移潜能呈负相关 (P <0 .0 1～ 0 .0 0 5) ,Bcl- 2和 Bax蛋白在 NSCL C中的表达呈负相关 (χ2 =9.93,P <0 .0 0 5)。结论　 Bcl- 2和 Bax蛋白相互抑制 ,均与 NSCLC发生、发展有关 ,可作为反映 NSCLC生物学行为的指标     

前列腺癌组织中血管形成的研究

目的　探讨前列腺癌 (PC)组织中微血管形成与其浸润、转移及预后的关系。方法　应用免疫组化SP法对 3 0例PC和 3 0例前列腺增生症 (BPH)组织进行CD3 4 表达并计数微血管(MV)。结果　PC组微血管密度 (MVD)显著高于BPH组 (P <0 .0 1 ) ,不同分级PC中也有显著性差异 (P <0 .0 1 ) ;浸润、转移性PC的MVD明显高于局限性PC组 (P <0 .0 1 ) ,2年内死亡者其MVD均高于存活 2年以上者 (P <0 .0 5 )。结论　肿瘤血管形成与PC分期、分级密切相关 ,癌细胞的浸润、转移有赖于肿瘤血管形成 ,MVD是反映前列腺癌生物学行为的重要指标。     

逆转录病毒介导CXCR4-siRNA对前列腺癌细胞体外侵袭能力的影响

目的设计构建趋化因子受体4(CXCR4)的RNA干扰逆转录病毒载体,探讨干扰CXCR4表达对前列腺癌细胞侵袭能力的影响。方法用PCR扩增CXCR4特异性小干扰RNA(smallinterferingRNA,siRNA),转入带有增强型绿色荧光蛋白(EGFP)和启动子U6的Pgensil-1质粒中,再将重组基因片段导入到逆转录病毒真核表达载体PLX-SN中,行限制性酶切及测序鉴定。重组逆转录病毒载体转染包装细胞PA317,获取病毒上清转染前列腺癌细胞,RT-PCR检测CXCR4mRNA表达。Transwell小室侵袭实验检测前列腺癌细胞体外侵袭能力的变化。结果通过酶切及测序鉴定,成功构建了PLXSN/EGFP-U6-siCXCR4。干扰质粒抑制CXCR4mRNA的表达,与空载体组比较,PC-3m、LNCaPsiRNA对CXCR4mRNA的抑制率分别为(84.26±10.20)%和(88.17±11.23)%。Transwell侵袭实验结果显示,干扰组微膜下孔细胞数PC-3m、LNCaP分别为(14.7±3.1)和(18.9±4.2)个,空载体组分别为(46.9±5.3)和(66.7±6.0)个,两者差异显著(P<0.05)。结论PLXSN/EGFP-U6-siCXCR4可以有效地抑制前列腺癌细胞CXCR4mRNA的表达,干扰CXCR4可以显著抑制前列腺癌细胞体外侵袭转移能力。     

应用免疫组织化学方法评价组织芯片的可靠性

目的探讨组织芯片技术在免疫组化试验中的可靠性和提高组织芯片可信性的方法。方法选择82例人乳腺癌标本石蜡蜡块,利用传统制片技术和自行研制的专用器具分别制作常规石蜡切片和组织芯片,采用免疫组化技术检测雌激素受体(ER)、孕激素受体(PR)的表达。结果采用自行研制的专用器具制备组织芯片,所得样本的可分析率均在92.9%以上,且随着对同一标本取材数量的增加,其可分析标本率也明显上升。采用传统方法检测的82例乳腺癌标本中ER和PR的阳性率分别为61%和58.5%;采用组织芯片技术,一式一份取材,其阳性率分别为51.9%和50.0%,一式两份方式取材,其阳性率分别为53.8%和53.8%,采用一式三份方式取材,其阳性率分别为57.1%和60.7%,且三种不同取材方式的检测结果与传统制片法之间并无统计学差异(P>0.75)。结论在采用统一制备标准并保证组织芯片质量的前提下,挖取直径1.5mm的组织样本制备组织芯片,尽管组织芯片技术与传统方法检测的结果的一致率仍随着对同一标本取材次数的增加而提高,但是三种不同取材方式的检测结果之间并无统计学差异。因此,一式一份取材制备组织芯片完全可替代传统制片技术用于免疫组织化学乃至原位杂交、荧光原位杂交技术的研究,且可保证组织芯片的可信性和高通量特征。     

REAL-TIME DETECTION OF SURVIVIN mRNA EXPRESSION IN CERVICAL CANCER CELL LINES USING MOLECULAR BEACON IMAGING

Objective To detect the expression of survivin mRNA in cervical cancer cell lines using molecular beacon imaging technology. Methods Human cervical cancer cells （HeLa and SiHa） and human fetal lung fibroblast HFL-I were cultured in vitro. After adding 100 nmol/L survivin mRNA molecular beacon, the fluorescent signals were observed under fluorescent microscope. The expressions of survivin in cervical cancer cells and HFL-I cell were examined by immunocytochemical streptravidin-biothin peroxidase （SP） assay at the same time. Results Two kinds of survivin mRNA molecular beacon, with different color fluorescence, had strong fluorescent signal in cervical cancer cell lines, and the signal in SiHa cell line was stronger, but these signals were not found in HFL-I ; Immunocytochemical staining of positive survivin was located in the cytoplasm of cervical cancer cell lines HeLa and SiHa, whereas, no expression of survivin was detected in HFL-I cell line. Conclusion The technology of molecular beacon imaging can be used to detect the expression of survivin mRNA in viable cells successfully, and may provide a new approach to the diagnosis of early stage cervical cancer and the following-up in the clinic.     

EXPRESSION AND SIGNIFICANCE OF BASIC FIBROBLAST GWOWTH FACTOR AND FIBROBLAST GROWTH FACTOR RECEPTOR-1 IN OVARIAN EPITHELIAL NEOPLASM

Basicfibroblastgrowedfactor (bFGF)wasakindofmultifunctionalgrowthfactorwhichhadahighaffcinitywithitsmainreceptorwhosenamewasfibroblastgrowthfactorreceptor 1(FGFR 1) .ByFGFRsuchasFGFR 1,bFGFcouldnotonlyin ductthegrowthofnewcapillary ,butalsostimulatethecell’smitokasisandpropagationaspowerfulmi tosigen ;Bystimulatingendothelialandtumorcelltosynthesizetheprofibrnolysinactivator(uPA ) ,re leasefibrinaseandcollagenase ,synthesizecollage nase ,sotoenhencetumorcell’sabilitytoinvadeentracell…     

人胃肠癌组织中浸润淋巴细胞活性的原位分析

利用一组抗淋巴细胞亚群的单克隆抗体和免疫组化方法对63例进展期胃癌和肠癌组织中浸润单个核细胞,尤其是散在浸润淋巴细胞(主要是T淋巴细胞)的I_2(HLA—DR)抗原和OKT_9抗原的表达进行了分析。同时,无选择的选取了18例慢性胃炎和慢性肠炎活检标本做为对照。在癌肿标本中,散在浸润淋巴细胞的I_2抗原表达显著变异。少数标本中很难查见I_2~+T细胞,大部分病例只能查见少数细胞呈I_2~+。63例中仅有10例其I_2~+细胞占全部散在浸润淋巴细胞的10—30%,所有瘤组织中几乎无OKT_9~+浸润T细胞。这种表现完全不同于正常粘膜及炎症粘膜中的I_2~+浸润单个核细胞分布。这说明就大多数病例来说,肿瘤组织中浸润的T细胞多数是非活化的。