结肠上皮良、恶性病变Ag-NOR及组织化学研究

应用银染核仁组成区(Ag-NOR)和 IBAS 全自动图象分析仪(西德:Kontron 公司)及组织化学技术,对结肠腺癌Ⅰ级56例(Ⅰ组)、Ⅱ级20例(Ⅱ组),结肠粘液腺癌15例(Ⅲ组),结肠腺癌之癌旁组织30例(Ⅳ组)和正常结肠组织30例(Ⅴ组)标本进行了形态定量和半定量研究。结果发现在上述各组细胞核中 Ag-NORs 颗粒平均数依次为6.14±1.70、7.73±3.50、8.09±2.71、3.90±0.17和2.73±0.42/核.癌的计数明显高于正常结肠和癌旁组织,有非常显著性差异(P<0.01)。Ag-NORs 颗粒的面积、等效圆直径结果相同,即为Ⅰ组>Ⅱ组>Ⅳ组>Ⅴ组>Ⅲ组;形状因子为Ⅲ组>Ⅱ组>Ⅰ组>Ⅳ组>Ⅴ组;周长为Ⅰ>Ⅳ组>Ⅱ组>Ⅲ组>Ⅴ组。上述各项参数指标作为结肠上皮良、恶性病变的鉴别和结肠腺癌的分类、分级的客观指标具有参考价值。组织学及组织化学染色在本实验中也具有重要参考意义。     

垂体腺瘤分类的免疫组织化学研究

目的：研究我国垂体腺瘤的功能分类及其与临床病理的联系。方法：采用Dako-Envision法进行免疫组织化学染色,检测45例手术切除垂体腺瘤中6种激素（GH,PRL,ACTH,TSH,LH,FSH）的表达情况,据此进行垂体腺瘤的分类及临床病理研究。结果：在45例垂体腺瘤中,生长激素细胞腺瘤为15例（33.3％）,催乳素细胞腺瘤6例（13.3％）,促肾上腺皮质激素细胞腺瘤9例（20％）,促性腺激素细胞腺瘤3例（6.7％）,多种激素细胞腺瘤4例（8.9％）,无功能细胞腺瘤8例（17.8％）。结论：垂体腺瘤中不同类型腺瘤所占的比例存在差异,生长激素腺瘤在外科切除的腺瘤中最常见。     

c-myc癌基因在大肠组织中表达的免疫组织化学和原位杂交研究

本文应用 c-myc 癌基因蛋白免疫组织化学和 mRNA 原位杂交方法研究了 c-myc 癌基因在大肠组织中的表达,并以胎盘作为阳性对照。发现在胎盘合体滋养叶细胞和结肠粘膜表面上皮都有 c-myc 癌基因的表达。结肠肿瘤 c-myc 表达增加,腺癌表达高于腺瘤。腺瘤中以绒毛状腺瘤表达最高。研究表明 c-myc 癌基因表达并不绝对与细胞增殖状态相关。     

结肠腺瘤发生的危险因素及和幽门螺杆菌感染的相关性分析

目的:分析结肠腺瘤发生的危险因素及和幽门螺杆菌(Hp)感染的相关性。方法:选择我院2018年6月～2018年12月收治的180例结肠腺瘤患者,同时选择我院接受结肠镜检查无异常者152例作为对照组。收集和比较两组患者的一般资料,采用14C尿氮呼气试验检测Hp的感染情况,多因素Logistic回归分析结肠腺瘤发生的危险因素。结果:多因素Logistic逐步回归分析结果显示男性、年龄、体质指数24 kg/m~2、腹型肥胖、饮酒、吸烟、喜食红肉、喜食果蔬、高脂血症、糖尿病、粪便隐血阳性、肿瘤家族史及Hp阳性是结肠腺瘤发生的危险因素,喜食果蔬为其发生的保护因素。HP阳性率组腺瘤1 cm、腺瘤数目多发、左结肠率高于Hp阴性组(P0.05);Hp阳性组和Hp阴性组肠腺瘤患者腺瘤蒂部分型、腺瘤病理类型比较差异无统计学意义(P0.05)。结论:结肠腺瘤的发生和多种危险因素有关,其中Hp感染可增加结肠腺瘤发生发展风险,临床应将此类高危人群作为结肠腺瘤的重点筛查对象,以降低结肠癌的潜在发生风险。     

实验性脾虚证大鼠结肠和盲肠组织化学、免疫组织化学研究

成年雄性Wistar大鼠60只,分正常对照组;脾虚组;自然恢复组和四君子汤治疗组。四组动物取结肠和盲肠分别进行H-E;酶组织化学和5-羟色胺细胞PAP免疫组织化学观察。结果表明,脾虚组结肠和盲肠粘膜上皮的杯状细胞较对照组明显增多,淋巴细胞及淋巴集结也增多。固有层毛细血管充血并有炎症细胞的增多。上皮细胞和腺细胞琥珀酸脱氢酶(SDH)减弱,乳酸脱氢酶(LDH)增强;结肠上皮细胞和腺细胞碱性磷酸酶(AIP)和三磷酸腺苷酶(ATPase)增强,而盲肠上皮细胞和腺细胞AIP和ATPase减弱。结肠和盲肠5-HT细胞免疫组化反应减弱。这些结果表明肠上皮细胞酶的变化和5-HT细胞分泌活性的变化与脾虚证的发生密切相关,可能是导致脾虚证原因之一。经四君子汤治疗后以上各项指标均比自然恢复组更接近于正常对照组,说明此药对消化道粘膜上皮酶活性和5-HT细胞分泌活性恢复正常有明显效果,从而起治疗作用。     

胃粘膜肠上皮化生SC3A的免疫组织化学研究

应用免疫组织化学ＡＢＣ方法等检测６２ 例胃良、恶性病变伴肠化组织中单克隆抗体ＳＣ３Ａ的表达及意义。结果： 癌旁肠化硫酸粘液阳性率明显较良性病变伴肠化高; 硫酸粘液阳性组肠化ＳＣ３Ａ阳性率明显高于硫酸粘液阴性组; 而且ＳＣ３Ａ 阳性率在酸性粘液阳性组胃癌明显高于酸性粘液阴性组; 硫酸粘液阳性组胃癌明显高于硫酸粘液阴性组。提示结肠型肠化可能与肠型胃癌的发生有一定关系     

促泌素在肾上腺原发肿瘤中的表达及其临床意义

目的比较促泌素(secretagogin,SCGN)与传统神经内分泌标记物在肾上腺原发肿瘤中的表达差异。方法收集肾上腺原发肿瘤手术标本共37例,其中包括18例皮质腺瘤、3例皮质腺癌、16例嗜铬细胞瘤。同时选取5例正常肾上腺组织,5例肾透明细胞癌作为对照。所有标本均使用SCGN、PGP9.5、CD56、NSE、Syn及CgA进行免疫组织化学SP法染色。结果SCGN在全部5例正常肾上腺皮质均有表达,而在髓质不表达(P<0.01),其中在皮质的表达明显高于PGP9.5和CgA的表达(P均<0.01);全部18例皮质腺瘤均表达SCGN,且明显高于NSE(P<0.05)、PGP9.5和CgA(P均<0.01);肾上腺嗜铬细胞瘤中SCGN的阳性表达率仅为18.8%(3/16),明显低于其它标记物(P均<0.01)。SCGN在皮质腺瘤(18/18)中的表达明显高于嗜铬细胞瘤(3/16)(P<0.01),而PGP9.5和CgA在嗜铬细胞瘤(15/16,16/16)中的表达明显高于皮质腺瘤(3/18,1/18)(P均<0.01);CD56、NSE和Syn在皮质腺瘤、皮质腺癌和嗜铬细胞瘤中均有高表达,但两两组间比较均无统计学差异(P均>0.05)。SCGN在全部5例肾透明细胞癌中均不表达。结论SCGN对肾上腺皮质腺瘤有较高敏感性,其与嗜铬细胞瘤的标记物CgA和PGP9.5联合可在两者的诊断和鉴别诊断中发挥重要作用。     

胃粘膜肠上皮化生与胃癌的粘液组织化学研究

应用粘液组化技术对１８５例胃良恶性疾病进行了研究。结果：胃良恶性疾病伴肠化检出率无明显差异;但含有硫酸粘液的结肠型肠化在胃癌癌旁检出率明显高于良性疾病组。不同分化程度胃癌的转移及术后生存率无明显差异;但含有酸性粘液的肠型、混合型胃癌的淋巴结转移率明显较仅含中性粘液的胃型胃癌高。提示：结肠型肠化与胃癌有一定关系;癌细胞产生大量酸性粘液可能有利癌细胞的扩散与转移,对胃癌病人的预后有一定影响     

脱钙对鼠颅骨标本中肥大细胞组织化学与免疫组织化学染色的影响

目的:探讨脱钙对鼠颅骨标本中肥大细胞组织化学与免疫组织化学染色影响.方法:用不同脱钙液处理小鼠颅骨标本,组织切片后进行苏木素一伊红染色、甲苯胺蓝染色、醛品红染色以及免疫组织化学染色.结果:经过不同脱钙液处理后的颅骨组织切片组织结构保存完好,肥大细胞的组织化学染色(甲苯胺蓝和醛品红染色),及肥大细胞中胰蛋白酶的免疫组织化学染色清晰.结论:不同脱钙液(8%盐酸脱钙液、EDTA脱钙液、混合酸脱钙液)处理不影响鼻腔粘膜中肥大细胞的组织化学和免疫组织化学染色.     

Histochemical reactions of gastrointestinal mucosubstances with orcein, high iron diamine and Alcian blue after prior oxidation of tissue sections.

The histochemical orcein reaction (orc) for mucosubstances in tissue samples from the human gastrointestinal tract was compared with PAS, high iron diamine (HID) and Alcian blue reactions at pH 1.0 or 2.5 (AB 1 and AB 2.5). Orc, HID and AB 1 reactions were performed also with prior oxidation of the tissue sections with potassium permanganate or performic acid (ox-orc, ox-HID and ox-AB reactions, respectively). Orc reaction stained mucosubstances similarly to HID and AB 1; only the brush border and goblet cells in the colon were stained. The reactions of the mucosubstances obtained with ox-orc differed from those with PAS, HID, AB 1 or AB 2.5 but were similar to those with ox-HID or ox-AB; the mucosubstances in the brush border and the goblet cells in the colon and small bowel and in the foveolar epithelium of the stomach were strongly stained. Pyloric and cardiac glands were stained faintly with ox-orc but not with ox-HID or ox-AB. Brunner's glands were negative with ox-orc, ox-HID and ox-AB reactions. It was assumed that the orc reaction stains, like HID or AB 1, sulphate groups in epithelial mucosubstances, and that sulphonic acid residues, resulting from oxidation of disulphide groups in the protein core of mucus glycoproteins, are responsible for the ox-orc as well as for the ox-HID and ox-AB reactions.     

The histochemistry of urea-unmasked glycosaminoglycans in the skin of the eel, Anguilla japonica

In the connective tissues of the dermis and subcutis of the eel skin, the histochemistry of urea-unmasked glycosaminoglycans has been studied by means of combined staining and enzyme digestion procedures. The staining procedures employed were alcian blue (AB) pH 1.0, AB pH 2.5, aldehyde fuchsin (AF), periodic acid-Schiff (PAS), AB pH 2.5-PAS, high iron diamine (HID) and low iron diamine (LID) methods, whereas the enzymes used were Streptomyces and testicular hyaluronidases, chondroitinases ABC and AC and keratanase. The results obtained have shown that a substantial amount of dermatan sulfate and a relatively small amount of hyaluronic acid, chondroitin, chondroitin sulfate A and/or C were the glycosaminoglycans involved in the connective tissues of the eel skin and that the tissues were devoid of keratan sulfate.     

Histochemical reactions of gastrointestinal mucosubstances with orcein,high iron diamine and alcian blue after prior oxidation of tissue sections

Summary The histochemical orcein reaction (orc) for mucosubstances in tissue samples from the human gastrointestinal tract was compared with PAS, high iron diamine (HID) and Alcian blue reactions at pH 1.0 or 2.5 (AB 1 and AB 2.5). Orc, HID and AB 1 reactions were performed also with prior oxidation of the tissue sections with potassium permanganate or performic acid (ox-orc, ox-HID and ox-AB reactions, respectively). Orc reaction stained mucosubstances similarly to HID and AB 1; only the brush border and goblet cells in the colon were stained. The reactions of the mucosubstances obtained with ox-orc differed from those with PAS, HID, AB 1 or AB 2.5 but were similar to those with ox-HID or ox-AB; the mucosubstances in the brush border and the goblet cells in the colon and small bowel and in the foveolar epithelium of the stomach were strongly stained. Pyloric and cardiac glands were stained faintly with ox-orc but not with ox-HID or ox-AB. Brunner's glands were negative with ox-orc, ox-HID and ox-AB reactions. It was assumed that the orc reaction stains, like HID or AB 1, sulphate groups in epithelial mucosubstances, and that sulphonic acid residues, resulting from oxidation of disulphide groups in the protein core of mucus glycoproteins, are responsible for the ox-orc as well as for the ox-HID and ox-AB reactions.The study was supported by grants from the Cancer Society of Finland, Foundation of Orion Corporation and from the Paulo's Foundation, Helsinki, Finland     

The histochemical specificity of High Iron Diamine—Alcian Blue

Summary The specificity of the High Iron Diamine—Alcian Blue pH2.5 (HID—AB 2.5) procedure was examined in tissue sites containing sialogycoproteins alone or differing proportions of sialo- and sulphosialoglycoproteins. Studies with HID in differing final concentrations of hydrochloric acid or sodium chloride confirmed that staining is dependent upon both the pH and the ionic strength of the dye bath and demonstrated a marked heterogeneity in the pKa of the anionic groups of sialosulphoglycoproteins. Use of the sequence High Iron Diamine—Alcian Blue pH 1.0 demonstrated that complete or almost complete staining ofO-sulphate esters occurred when HID was prepared in water (final pH 1.3). However, under these conditions HID—AB 2.5 was shown to be non-specific because only black HID staining was observed in sites containing large quantities of sialic acids. This non-specificity was due either to the masking of Alcian Blue staining by HID and/or the black HID staining of anionic groups other than sulphate. These results may account for some of the conflicting data obtained by different groups of investigators who have studied transitional mucosa in human colonic diseases. Caution should be used in drawing conclusions from the use of HID—AB 2.5 without confirmatory evidence from other more specific procedures.     

CDX2在胃癌及癌前病变组织中的表达及其临床意义

目的研究CDX2在胃癌及癌前病变组织中的表达及其与临床病理因素之间的关系。方法采用免疫组织化学S-P法检测CDX2在胃癌及癌前病变组织中的表达,采用阿新兰/过碘酸雪夫氏染色(AB/PAS)和高铁二铵/阿新兰(HID/AB)染色对胃黏膜肠化生进行分型。结果 (1)CDX2在正常胃黏膜组织、慢性浅表性胃炎中不表达,CDX2在肠化、异型增生及胃癌中的阳性率(81.7%、50%、48.2%)均明显高于正常胃黏膜组织(P0.05),在肠化组织中的表达明显高于异型增生和胃癌(P0.05),在异型增生和胃癌中的阳性率差异无显著性(P0.05)。CDX2的表达与肠化的分型和异型增生的分级之间均无关系。(2)CDX2在肠型胃癌中的阳性率(67.4%)明显高于弥漫型和混合型胃癌(25.7%、42.9%);CDX2的表达与胃癌分化程度呈正相关,而与淋巴结转移和临床分期呈负相关,差异均有统计学意义。CDX2阳性表达组的五年生存率明显高于阴性表达组(P=0.038);多种变量对胃癌患者术后生存时间影响的分析显示,CDX2的表达和淋巴结转移是影响胃癌预后的独立因素(P0.05)。结论 CDX2可能在胃粘膜肠化生的发生及胃癌的发生发展中起重要作用,可作为胃癌诊断、判断胃癌的恶性程度及患者预后的有用指标。     

Histochemical analysis of extracellular matrix material in embryonic trisomy 1 mouse eye

Trisomic animals produced from mice doubly heterozygous for Robertsonian translocation chromosomes [Rb(1.3)/Rb(1.10)]consistently show eye defects (e.g., aphakia, microphakia, and retention of lens stalk). To determine if changes in distribution or composition of extracellular matrix material may be a factor in development of these defects, eye structures of trisomy (ts) 1 embryos and normal littermates were studied histochemically using the following methods: Alcian blue 8GX, pH 2.5; periodic acid-Schiff (PAS), Alcian blue/PAS combined; high-iron diamine (HID), and HID/Alcian blue combined. Eye development was divided into stages to account for the known delay in ts 1 mouse development. Differences were found in staining patterns as early as stage 1. In later stages, the most consistent difference was an increased period of contact between lens and optic cup due to retardation of interface matrix dissolution between these rudiments in ts 1 embryos. Eyes in which this occurred had abnormally shaped lenses. Overall, the ts 1 optic cup appeared to have fewer staining abnormalities and dysmorphology than did the lens or interface matrix. Triplication of a chromosome may indirectly alter temporal and spatial organization of extracellular matrix through action on cells responsible for the production of this material. Possible mechanisms of action are discussed.     

Fascin在结肠癌中表达的临床病理学研究

目的:Fascin是一种肌动蛋白结合蛋白,在多种上皮性肿瘤中高表达并与肿瘤侵袭有关。在本研究中观察fascin在结肠癌中表达及探讨其临床病理意义,为其在结肠癌早期诊断和预后预测方面的应用提供依据。方法:应用免疫组织化学技术检测Fascin在结肠肿瘤中的表达,并分析其表达的结肠癌临床病理意义。结果:Fascin在癌旁肠粘膜、腺瘤、腺癌中表达有显著性差异,其中癌旁肠粘膜组和腺瘤组阳性表达率无显著性差异(X2=0.344,P0.05),腺癌组阳性表达率显著高于癌旁粘膜组(X2=8.492,P0.0,1),腺癌组阳性表达率显著高于腺瘤组(X2=7.450,P0.01)。Fascin表达与结肠癌患者的年龄、性别、肿瘤浸润肠壁深度、淋巴结转移、分化程度无显著相关性,而在中-晚期病例(III/IV期)中的表达率显著高于早期病例(P0.05)。Fascin表达阳性病例的生存期显著高于Fascin表达阴性病例(P0.022)。结论:Fascin表达与结肠癌发生和预后密切相关,可能作为结肠癌早期诊断和预后的标志物。     

Seasonal variation in glycoconjugates of the pedal glandular system of the rayed Mediterranean limpet,Patella caerulea (Gastropoda: Patellidae)

Glycoconjugates secreted by the pedal system of the rayed limpet, Patella caerulea, were characterised in situ by histochemical and lectin-histochemical methods in individuals collected around the annual cycle, in November, March, and June. Stainings with periodic acid–Schiff (PAS), Alcian blue pH 2.5 (AB pH 2.5), Alcian blue pH 1.0 (AB pH 1.0), high-iron diamine–Alcian blue pH 2.5 and lectin binding assays with 9 lectins (Con A, WGA, succinylated-WGA, PNA, DBA, SBA, AAA, UEA-I, LTA) were performed. Four secreting cell types were observed in the sole, one in the peripheric region, and two in the sidewall. Glycoconjugate composition varied among cell types and also in one and the same cell type throughout the year. β-Elimination followed by PAS and AB pH 2.5 stainings indicated that most saccharidic chains were O-linked to the protein backbone. Secretion by sole and peripheric region was acidic, carboxylated and/or sulfated, whereas that of the sidewall was neutral. Glucosaminylated and 1,4-fucosylated residuals were predominant in the cell types along the year, 1,2-fucosylated residuals being observed only in the sidewall cells in June. Mannosylated and/or glycosylated residuals were observed in all cells mostly in November. Galactosylated/galactosaminylated residuals were present mostly in the sidewall cells and in the sole subepidermal mucocytes in June. Mannosylated and/or glycosylated residuals in November are probably linked to gonad maturation or to higher locomotion and foraging activity, whereas galactosaminylation in the sole cells and 1,2-fucosylation and glucosaminylation in the sidewall cells in June are linked to a prolonged stationary state, increasing water adsorption to counteract dehydration and/or to modulate microbial interactions.