胚泡表面Le~Y寡糖对小鼠胚泡纤维粘连蛋白、层粘连蛋白表达及分泌的影响

目的　体外观察 L e Y 寡糖对着床前小鼠胚泡细胞外基质 ( ECM)主要成分纤维粘连蛋白 ( FN)、层粘连蛋白 ( LN)的表达和分泌的影响。　方法　采用特异性单克隆抗体 AH6阻断胚泡表面 L e Y寡糖 ,运用半定量逆转录 -聚合酶链反应 ( RT-PCR)及免疫印迹法检测 L e Y寡糖对着床前小鼠胚泡 FN及 LN的表达和分泌的影响。　结果　在体外培养中 ,经 AH6封闭胚泡表面 L e Y 寡糖抗原后仅 1 .5h,胚泡 FN及 L N的分泌有明显升高 ( P<0 .0 1 ) ,并且持续 6 h以上 ,而胚泡 FN及 L N的基因转录表达变化不明显 ( P>0 .0 5)。　结论　胚泡表面的 L e Y寡糖抗原对着床前胚泡的 FN及 L N的分泌具有促进作用     

骨桥蛋白在人正常月经周期子宫内膜的表达

目的研究骨桥蛋白(osteopontin,OPN)在人正常月经周期子宫内膜的表达。方法采用免疫组织化学法对51例人正常月经周期子宫内膜OPN蛋白质进行定位和定量检测,Western blotting法测定增殖期及分泌期OPN蛋白水平,采用逆转录-聚合酶链反应(RT-PCR)方法检测18例人正常月经周期子宫内膜增殖期及分泌期OPN mRNA的表达。结果OPN蛋白质主要分布在正常子宫内膜腺上皮细胞,增殖早期、中期无表达,增殖晚期出现,分泌中期、晚期表达最强。月经期子宫内膜腺上皮细胞表达较强。OPN mRNA表达分泌期强于增殖期(P<0.05)。Western blotting检测结果显示正常子宫内膜组织中存在OPN蛋白,分泌期含量较高。结论OPN蛋白质及mRNA在正常子宫内膜中的表达呈现明显周期依赖性。分泌中、晚期表达增强提示其可能参与胚胎着床。     

纤维连接蛋白、层粘连蛋白、胶原蛋白Ⅳ在大鼠膀胱癌组织的表达

目的 观察纤维连接蛋白(FN)、层粘连蛋白(LN)和胶原蛋白Ⅳ(COL Ⅳ)在大鼠膀胱癌组织的表达.方法 选取正常SD大鼠膀胱组织及大鼠原位膀胱癌模型建立后的非肌层浸润性膀胱癌(NMIBC)组织标本(每组20例),用免疫荧光及测定单位面积荧光强度(F/S)的方法比较FN、LN和COLⅣ表达的变化.结果 FN和LN在NMIBC的表达与正常大鼠膀胱组织比较,F/S值分别为FN(6.91 ±1.39和0.82±0.22)、LN(6.57±2.39和0.90±0.46),差异有统计学意义(P＜0.05);COLⅣ在NMIBC和正常大鼠膀胱组织F/S值为1.05±0.57和0.84±0.37,差异无统计学意义(P＞0.05).结论 FN和LN在NMIBC表达明显升高,COL Ⅳ的表达差异无统计学意义.     

IL-18及IL-18结合蛋白在人正常月经周期子宫内膜的表达

目的探讨白细胞介素(IL)18及IL18结合蛋白(IL18BP)在人正常月经周期子宫内膜上的表达。方法取因子宫肌瘤或子宫脱垂行全子宫切除术的患者子宫内膜,按月经周期和子宫内膜形态学检查结果分为增生期和分泌期2组,每组10例。免疫组织化学法检测子宫内膜的IL-18及IL-18BP表达;实时定量聚合酶连锁反应(qRT-PCR)检测子宫内膜IL-18及IL-18BP mRNA的表达。结果 IL-18、IL-18BP在人增生期和分泌期子宫内膜中均有表达,且随着月经周期其表达呈现时空性变化。IL18在增生期表达于腔上皮、腺上皮细胞膜及基质细胞,与增生期相比,分泌期基质细胞IL18的阳性表达显著降低(MOD值0.33±0.07 vs.0.52±0.12,P0.05),而IL18BP的表达则显著增加(MOD值0.33±0.20 vs.0.10±0.07,P0.05)。qRT-PCR结果显示,在增生期和分泌期子宫内膜标本均有IL18和IL18BP mRNA表达,与增生期相比,分泌期IL18表达显著下降(P0.05),IL18BP表达显著增强(P0.05),IL18BP/IL18比值显著增高(P0.05)。结论 IL18和IL18BP在整个月经周期人子宫内膜上均有表达,且其表达呈现时空性的变化,可能与月经周期过程中子宫内膜的崩解修复有关。     

微血管密度和层粘连蛋白在直肠癌组织中的表达

基底膜是宿主与癌细胞的重要屏障,层粘连蛋白(laminin,LN)是基底膜的重要组成成份.微血管密度(microvessel density,MVD)或LN作为肿瘤的预后指标已有报道[1].为了研究MVD与LN在直肠癌组织中的表达特点及其相互关系,我们检测了31例直肠癌MVD和LN.     

低温保存皮肤纤维连接蛋白和层粘连蛋白的表达

目的:观察细胞外基质中纤维连接蛋白(FN)、层粘连蛋白(LN)在不同温度保存皮肤组织中的分布,以探讨其与皮肤冷冻损伤的关系.方法:用FN、LN抗体对5种保存条件下的皮肤作免疫组化染色,用图像分析仪测定表皮组织中的含量,并用H.E染色观察皮肤的组织结构变化.结果:与新鲜皮片组相比,-196℃H抗冻液中保存皮肤组织结构保存较好,FN、LN含量变化不明显,而4℃、-20℃、-80℃保存皮肤组织结构破坏明显,含量亦明显下降.结论:皮肤低温保存冷冻损伤与细胞外基质中FN、LN破坏有关.     

CD44和纤粘连蛋白在子宫腺肌病中的表达及其意义

目的探索CD44和纤粘连蛋白因子(FN)在子宫腺肌病(AMS)患者子宫内膜中的表达情况,评估其在AMS中的潜在诊断价值。方法采用免疫荧光染色鉴定子宫内膜细胞中CD44和FN的表达和细胞定位,收集AMS患者和对照组子宫内膜进行体外分离培养,分别提取mRNA和蛋白质。采用反转录定量PCR(RT-qPCR)检测CD44和FN的mRNA表达情况,利用Western blot检测CD44和FN的蛋白表达水平,并进行组间比较。结果 CD44的mRNA和蛋白相对表达量在两组患者中均无显著性差异(P0.05);与对照组比较,FN的mRNA相对表达量在AMS患者子宫内膜中呈现约6.5倍的下调,蛋白质表达呈现约1.8倍的下调,差异均具有统计学意义(P0.01)。结论 FN参与AMS病理过程中细胞粘附生长的调节,FN的表达下调以及CD44的稳定表达有望作为AMS患者诊断或者预后的潜在预测指标。     

丹蒲胶囊对试验性慢性前列腺炎的病理学和免疫组织化学研究

目的 探讨丹蒲胶囊治疗慢性前列腺炎的作用机理。方法 对实验性大鼠前列腺组织进行组织病理学和纤维连接蛋白（FN）、层粘连蛋白（LN）免疫组织化学的观察。结果 丹蒲胶囊可明显抑制大鼠慢性前列腺炎组织中的炎细胞浸润和间质纤维组织增生，并促使前列腺腺腔扩大，腔内分泌物增多，丹蒲胶囊组与模型组相比，有显著性差异（P＜0．05）。丹蒲胶囊组与模型组的FN表达均呈弱阳性，两组相比，无显著性差异（P＞0．05）；丹蒲胶囊组的LN表达主要呈弱阳性，模型组的LN表达呈强阳性和阳性，两组相比，有显著性差异（P＜0．05）。结论 丹蒲胶囊具有明显抗炎、抗纤维化及增强腺细胞的分泌功能。FN、LN在前列腺炎发病中起重要作用，LN可作为判断慢性前列腺炎的炎症及纤维化程度的参考指标，丹蒲胶囊可显著降低大鼠慢性前列腺炎组织内的LN水平。     

胰岛素样生长因子结合蛋白1-3在人早孕期蜕膜和绒毛中的表达及其作用

目的探讨胰岛素样生长因子结合蛋白1-3(IGFBP1-3)在早孕期蜕膜和绒毛中的表达及其作用。方法选取早孕5~7周行人工流产术的蜕膜和绒毛标本共48例,每一孕周分早、中、晚三个时期,每个时期5~6例,以分泌中期(周期第21天)子宫内膜标本为对照。采用免疫组织化学LsAB法和计算机图像分析法对IGFBP1-3在围着床期子宫内膜、蜕膜和绒毛滋养细胞的表达进行定位和定量分析。结果IGFBP1-3在子宫内膜腺、腔上皮、蜕膜细胞和绒毛滋养层中都有表达,其表达在孕5~6周达峰值,与对照的分泌中期内膜比较,差异有统计学意义(P<0.01)。结论IGFBP1-3在早孕期表达增强,推测可能参与了子宫内膜的蜕膜化、滋养层细胞侵袭性以及早期胚胎发育的调节过程。     

正常月经周期子宫内膜瘦素和瘦素受体的表达

目的　探讨瘦素和瘦素受体系统在正常月经周期子宫内膜表达特点及其意义。　方法　应用免疫组织化学方法检测 60例正常月经周期子宫内膜瘦素和瘦素长受体蛋白的表达。原位杂交技术测定 2 5例子宫内膜瘦素和瘦素长受体mRNA。　结果　瘦素蛋白在子宫内膜腺体和间质呈阳性或强阳性表达 ,月经周期各期间无显著差异 ;瘦素长受体蛋白在子宫内膜间质中呈弱阳性或阴性表达 ,月经周期各期间无显著差异 ;瘦素长受体蛋白在子宫内膜腺体的表达分泌期显著高于增殖期 (P <0 .0 0 1 )。瘦素和瘦素长受体mRNA在增殖期和分泌期子宫内膜的腺体和间质中均呈阳性表达。　结论　瘦素长受体蛋白在分泌期子宫内膜腺体表达增强 ,有可能在孕卵着床过程中发挥作用     

Expression of vascular endothelial growth factor and its two receptors in normal human endometrium

<正> Objectives:We try to demonstrate the expression of vascular endothelial growthfactor (VEGF) and its receptors,flt-1 and KDR,in normal human emdometrium duringthe menstrual cycle.Methods:Immunohistochemical method was used to observe the expression ofVEGF and its two receptors in emdometrium throughout the normal menstrual cyclemeanwhile the isoforms of VEGF were also detected by Western blot analysis.The en-dothelial cells of micro-vessels were marked with VIII factor antibody.Results:VEGF and its receptors existed in endometrial glandular,stromal and vas-cular endothelial cells of human endometrium.Their expressions were higher in the mid-secretory phase of menstrual cycle and highest at menstruation.VEGF_(121) and VEGF_(165)were the predominant isoforms in normal human endometrium.Conclusion:The expression of VEGF and its two receptors showed cycle-dependentin human endometrium,probably involved in embryonic implantation and endometrialproliferation and differentiation.     

无排卵性功能失调性子宫出血患者子宫内膜雌、孕激素受体及基质金属蛋白酶表达变化的研究

目的研究无排卵性功能失调性子宫出血(功血)患者子宫内膜雌激素受体(ER)、孕激素受体(PR)、IV型胶原、基质金属蛋白酶-9(MMP-9)及其抑制物-1(TIMP-1)的表达,探讨无排卵性功血的发病机制。方法采用免疫组化法检测20例正常增殖期子宫内膜(对照组)和66例无排卵性功血患者(病例组)子宫内膜腺上皮和基质细胞ER、PR、IV型胶原、MMP-9及TIMP-1表达。结果病例组子宫内膜腺上皮和基质ER、PR表达高于对照组(P<0.05),MMP-9水平明显增加(P<0.05),MMP-9/TIMP-1比值升高(P<0.01),IV型胶原含量明显下降(P<0.01)。相关分析显示,无排卵性功血患者子宫内膜MMP-9水平与ER、PR均呈正相关(r=0.605,P<0.05;r=0.697,P<0.05)。结论无排卵性功血患者子宫内膜出血原因可能与雌激素作用下ER、PR表达失调有关,且MMP-9水平异常升高,导致细胞外基质过度降解,临床表现为子宫不规则出血。     

A clinicopathological study of the expression of extracellular matrix components in urothelial carcinoma

OBJECTIVE: To measure the immunohistochemical expression of the extracellular matrix (ECM) components tenascin, fibronectin, collagen type IV and laminin in urothelial carcinomas, and to correlate their expression with clinicopathological features to clarify the prognostic value of these molecules and their role in tumour progression. MATERIALS AND METHODS: Tumour specimens obtained during transurethral resection of bladder tumour (TURBT) from 103 patients (82 men and 2 1 women, mean age 66.7 years, range 27-89) were studied retrospectively. The expression of tenascin, fibronectin, collagen type IV and laminin was correlated with clinicopathological features (tumour grade and stage, multiplicity, simultaneous in situ component, the proliferative activity as estimated by the two proliferation associated indices, Ki-67 and proliferating cell nuclear antigen, the recurrence rate, and the progression of invading tumour). Specimens investigated for tenascin expression from patients with superficial bladder cancers were categorized into 28 treated by TURBT only and 53 who had TURBT followed by intravesical instillations of interferon. RESULTS: Cytoplasmic tenascin expression was detected in tumour cells in 20% of specimens. Tenascin was expressed in the tumour stroma in 76% of specimens, and was positively correlated with tumour grade and stage. Stromal tenascin expression was positively correlated with proliferative activity, and with the expression of fibronectin and collagen type IV. Fibronectin was expressed in the tumour stroma in 89% of specimens and was positively correlated with tumour stage, proliferative activity, and expression of collagen type IV and laminin. Collagen type IV was expressed in 93% of specimens, and was positively correlated with tumour grade and stage. Laminin was expressed in 78% of specimens and had no significant correlation with the clinicopathological features. Patients treated with TURBT alone and who had low levels of tenascin had a longer tumour-free interval than those with high levels of tenascin. CONCLUSION: Levels of tenascin might be valuable for predicting the risk of early recurrence. The expression of tenascin, fibronectin and collagen type IV seems to be correlated with more aggressive tumour behaviour. Furthermore, their interrelationships could indicate that they are involved in the remodelling of bladder cancer tissue, probably influencing tumour progression.     

HOXA11在人子宫内膜和早孕蜕膜中的表达及与不育的关系

目的检测HOXA11蛋白在正常月经周期子宫内膜、早孕蜕膜和不明原因不育子宫内膜组织中的表达,探讨HOXA11与不明原因不育及雌孕激素受体(ER、PR)表达的相关性。方法采用免疫组织化学和Western-Blot方法对38例正常子宫内膜、15例早孕蜕膜和19例不明原因不育子宫内膜组织中HOXA11、ER及PR的表达。结果HOXA11蛋白在子宫内膜腺上皮和基质细胞均有表达,以分泌中晚期子宫内膜和早孕蜕膜表达量较高;在不育内膜的表达量显著降低(P<0.05);ER、PR在各组表达量的变化与HOXA11相同,即分泌中晚期子宫内膜和早孕蜕膜的表达量较高,而在不育内膜的表达量显著下降(P<0.05)。结论HOXA11基因在着床期子宫内膜的分化、发育以及着床后妊娠的维持起一定作用;HOXA11表达量下降与ER、PR表达量降低相关;HOXA11表达下降或缺失可能是女性不明原因不育的原因之一。     

Expression of basement membrane components in odontogenic tumors.

OBJECTIVE: The objective was to characterize the expression of BMCs (laminins 1 and 5, collagen type IV, and fibronectin) in ameloblastomas, calcifying cystic odontogenic tumors (CCOTs), and adenomatoid odontogenic tumors (AOTs). STUDY DESIGN: BMCs were analyzed in 14 ameloblastomas, 7 CCOTs, and 7 AOTs using immunohistochemistry. RESULTS: In normal oral mucosa, linear deposits of these proteins were found at the epithelial-mesenchymal junction, but not in epithelial cytoplasm. In all tumors studied, linear deposits of all proteins were found at the epithelial-mesenchymal junction; laminin 1 was expressed in all tumor cells, regardless of cell types. For CCOTs, laminin 5 was found faintly in suprabasal cells, but expressed strongly in ghost cells. For AOTs, laminin 5 strongly decorated tumor cells adjacent to mineralization. CONCLUSIONS: Laminin 1 may be a marker for odontogenic epithelium. Additionally, laminin 5 may be involved in ghost cell formation and initiation of calcification.     

Comparison of extracellular matrix antigens in subtypes of bronchioloalveolar carcinoma and conventional pulmonary adenocarcinoma. An immunohistochemical study.

In contrast to the conventional pulmonary adenocarcinomas (CPAs), bronchioloalveolar carcinoma (BAC) grows predominantly by spreading along the existing alveolar septal framework. Within the BAC category, three subtypes have been identified: mucinous, nonmucinous, and sclerosing BAC. Of these, mucinous and sclerosing BACs have worse prognoses compared with nonmucinous BAC. However, the manifestation of aggressive behavior is different between the mucinous and sclerosing types of BACs. Multifocality is often produced by aerogenous spread, especially in the case of mucinous BACs. To study the differences between the BAC subtypes and the conventional pulmonary adenocarcinomas, we employed a battery of immunohistochemical stains marking the extracellular matrix architecture (laminin, collagen IV, fibronectin, and collagen III), a degradative enzyme against a basement membrane component (anti-type IV collagenase) and cellular receptors for laminin and collagen IV (alpha 2 integrin) on 16 BACs (5 mucinous, 5 nonmucinous, and 6 sclerosing) and 30 CPAs. The mucinous and nonmucinous BACs demonstrated neoplastic epithelial cells growing along a continuous basement membrane. A similar growth pattern with intact basement membrane was noted in the periphery of sclerosing BACs. However, in contrast to mucinous and nonmucinous BACs, all cases of sclerosing BACs showed disruption or complete absence of basement membrane components (laminin and collagen IV) around the embedded glands located centrally in the sclerotic fibrous stroma, as was seen in the basement membrane analysis of conventional adenocarcinomas. Furthermore, increased type IV collagenase activity was seen in the small centrally located embedded glands in comparison to the peripheral glands. These architectural alterations of basement membrane disruption and phenotypic expression of degradative activity may be a reflection of the invasive behavior of the sclerosing BACs and their tendency to produce lymph node metastasis. Although the mucinous BACs did not show evidence of basement membrane disruption, there was a marked increase in their levels of type IV collagenase expression along with consistently low levels of alpha 2 integrin receptor (laminin and collagen IV receptor) expression. These findings may be related to the ability of the mucinous BACs to detach from the underlying basement membrane and spread aerogenously, and is to be contrasted with the stromal infiltration and desmoplasia of sclerosing BACs and CPAs.     

Increased and localized accumulation of chondroitin sulphate proteoglycans in the hyperplastic human prostate

OBJECTIVE: To analyse by immunohistochemistry the expression of chondroitin sulphate (CS) (detected in the hyperplastic prostate and possibly affecting the proliferation of prostate cells) in benign prostatic hyperplasia (BPH) to determine its distribution and location. MATERIALS AND METHODS: Samples of BPH were obtained from 11 patients (aged 58-83 years) and controls consisting of the transitional zone of five prostates from young men aged 19-27 years. Tissue sections were labelled with antibodies against CS, perlecan, type IV collagen, laminin, and smooth muscle cell (SMC) alpha-actin. The amount of CS immunostaining was estimated by semi-quantitative scoring and correlated with prostate-specific antigen (PSA) level and prostate size. RESULTS: The anti-CS antibody faintly stained the stroma of normal prostates, but in BPH samples the staining was intense and concentrated around acini, including the periphery of adjacent SMCs. This staining pattern was totally absent in the normal samples. Type IV collagen, perlecan and laminin were homogeneously distributed in the whole stroma of both normal and BPH samples. There was no significant correlation between intensity of CS staining and either PSA or prostate size. CONCLUSIONS: The expression of CS proteoglycans is increased in BPH, where they co-locate with basement membranes of the acinar epithelium and of peri-acinar SMCs. This enhanced expression is specific for these proteoglycans, as other basement membrane components are unaffected, and this may result from the regulatory effects of local factors that are active in BPH.     

Increased matrix gene expression by glucose in rat neural connective tissue cells in culture

The effects of different glucose concentrations on the expression of extracellular matrix genes were examined in primary cell cultures initiated from sciatic nerves of nondiabetic Sprague-Dawley rats. The cells were incubated in medium containing 5.5, 15, or 25 mM D-glucose, and the expression of type I and IV collagens, laminin, and fibronectin genes was examined at mRNA steady-state levels by Northern hybridizations. Incubation of cell cultures, consisting of Schwann cells, perineurial cells, and fibroblasts, in high glucose concentrations (15 or 25 mM D-glucose) resulted in elevation of pro-alpha 1(IV), pro-alpha 2(IV), and pro-alpha 1(I) collagen chain and fibronectin mRNAs after only 3 days of incubation, whereas laminin B2 chain mRNA levels appeared unaltered. These observations suggest that hyperglycemia may partially contribute to basement membrane thickening in peripheral nerves of diabetic individuals by increasing the expression of genes coding for basement membrane components, e.g., type IV collagen.     

Fibronectin, laminin, and collagen types I, III, IV and V in the healing rat colon anastomosis.

The purpose of this work was to study normal anastomotic healing in the rat colon. The unprepared sigmoid colon was divided and a colo-colostomy performed using a one-layer inverting technique. Frozen sections were taken and studied immunohistologically with specific antibodies to fibronectin, laminin and collagen types I, III, IV and V. From day one onwards a strong fibronectin reaction was observed in the anastomosis, reaching maximum staining intensity on postoperative day five. Type III collagen and pericellular type V collagen were at first detected in the anastomosis on day two. From day three onwards all collagens studied and laminin were present in the repair tissue (laminin and type IV and V collagen in the regenerating capillary walls). Maximum immunofluorescence was observed on day seven and it remained on a high level throughout the study, except for fibronectin, which weakened gradually after the fifth postoperative day. The results indicate that healing of the colon anastomosis occurs by rapid accumulation of connective tissue components between the inverted leaves of the colonic wall, as also new capillaries consisting of the basement membrane components, type IV and V collagen as well as laminin, are formed.