CD45RO~+T细胞和CD68~+细胞在外阴上皮内非瘤样病变和外阴癌中的侵润

目的：探讨CD45RO^＋T细胞和CD68^＋细胞在外阴上皮内非瘤样病变和外阴癌发生、发展过程中的侵润及作用。方法：免疫组化S-P法分别检测CD45RO^＋T细胞、CD68^＋细胞在各20例外阴鳞状上皮细胞增生、外阴硬化性苔藓、外阴鳞状上皮癌病变中的侵润,取10例外阴正常皮肤作对照。结果：CD45RO^＋T细胞和CD68^＋细胞在外阴正常皮肤中侵润水平极低,明显低于外阴鳞状上皮细胞增生、外阴硬化性苔藓和外阴鳞状细胞癌病变（P〈0.05）。它们在外阴硬化性苔藓晚期病变中侵润明显高于早期病变（P〈0.05）;在中高分化外阴鳞状细胞癌组织中侵润明显高于低分化外阴鳞状细胞癌组织（P〈0.05）。CD45RO^＋T细胞在中高分化外阴鳞状细胞癌组织中侵润最高,其次是晚期硬化性苔藓（P〈0.05）;CD68^＋细胞在中高分化外阴鳞状细胞癌和晚期硬化性苔藓病变中侵润最高,但二者之间没有统计学差异（P〉0.05）;它们均明显高于外阴鳞状细胞增生、低分化外阴鳞状细胞癌和早期硬化性苔藓（P〈0.05）,而在后三者之间侵润没有明显差异（P〉0.05）。CD68^＋细胞和CD45RO^＋T细胞在外阴不同病变中侵润具有正相关性,相关系数为0.742（P〈0.001）,而且CD68^＋细胞多出现在CD45RO＋T细胞的周围,且多在病变浅层。结论：CD45RO^＋T细胞和CD68^＋细胞参与皮肤的免疫反应,它们相互协同作用,在外阴上皮内非瘤样病变和外阴癌发生发展过程中发挥免疫调节作用。     

外阴营养不良患者病变组织中性激素受体的检测

目的：研究外阴营养不良与性激素受体的关系。方法：采用直接荧光法，测定了１１例患者病变组织的性激素受体（ＳＨＲ）。其中增生型４例，硬化苔藓型４例，混合型３例。并对其中８例进行了外阴未发病组织ＳＨＲ对照。结果：（１）患者病变组织及未发病组织表皮各层及真皮层内均有不同程度的雌激素受体（ＥＲ）、孕激素受体（ＰＲ）、雄激素受体（ＡＲ）阳性率；（２）患者病变组织及未发病组织基底层ＥＲ、ＡＲ无或少于表皮其它各层     

角化型外阴鳞状细胞癌中p16和cyclin D1蛋白的表达

外阴鳞状细胞癌（vulvar squamous cell carcinomas,VSCCs）多发生于老年妇女,但也见于年轻的患者。最近一些病理学家提出VSCCs的病因可能分为两类：一类与老年妇女外阴上皮内非瘤样病变（NNED）有关,如硬化苔癣（LS）、鳞状上皮增生（SCH）等;另一类多发生于年轻的妇女,与人乳头瘤病毒（HPV）感染有关。但LS和SCH与VSCCs关系尚不明确。有关p16、cyclin D1的改变在外阴癌中的作用研究报道甚少,[第一段]     

人胚食管上皮凝集素结合部位的观察

选用四种凝集素(WGA、UEA、LCA、BSA)对人胚食管采用ABC法进行石蜡切片的标记.结果发现四种凝集素对不同胚龄的食管呈不同反应.LCA在8月开始于基底层细胞出现微弱阳性反应.在3月,WGA巳能与基底层细胞膜明显结合,UEA和BSA可与非纤毛区细胞结合,而不与纤毛区细胞结合.出生时,四种凝集素的染色均未达到成人食管的染色强度.本研究结果提示,上述凝集素在人胚食管上皮的结合部位的分布情况及染色强度,可作为该上皮细胞的发育、分化程度的标志之一.     

比较研究人正常,增生性胃粘膜和胃癌时对酶标凝集素的反应

本文用酶标凝集类,对35例胃手术病人的正常胃粘膜、增生性胃粘膜、小肠化生和胃癌等与凝集素的反应,在光镜上作了比较观察。结果表明正常胃粘膜的上皮细胞对PNA、DBA和ConA均为阴性。而胃小凹上皮PNA和DBA则为阳性。增生肥大的胃粘膜、上皮细胞和胃小凹上皮对PNA和DBA均匀阳性反应,其染色强度在胃小凹上皮有所增强。化生的胃粘膜,PNA阳性,但DBA阴性。胃腺的主细胞仅对ConA染色。壁细胞能选择性地与DBA结合,有的壁细胞也能与PNA结合。癌细胞与凝集素反应的能力比增生性粘膜减弱,可能是由于癌细胞缺少糖苷化作用酶的关系,     

甲胎蛋白异质体的检测及临床应用

甲胎蛋白（alpha fatoprotein，AFP）是早期肝细胞癌（HCC）诊断的良好指标，但是部分良性肝病也有AFP升高，30％～40％的肝细胞癌AFP正常，这样导致部分肝病无法判断良恶性。研究发现，AFP存在糖链有差别的异质体，并利用它与不同植物凝集素的亲和进行分类，有小扁豆凝集素（lens culinaris lectin，LCA）型和刀豆素A（concanavalin A，ConA）型等。据AFP和LCA亲和的强弱程度分为AFP—L1、AFP-L2、AFP-L3，AFP—L1是LCA非结合型，     

分化型外阴上皮内瘤变的研究进展

外阴鳞状细胞癌约占外阴恶性肿瘤的80%, 高级别外阴鳞状上皮内瘤变是外阴鳞状细胞癌的癌前病变, 按照病理机制可分为普通型高级别外阴鳞状上皮内瘤变, 与人乳头状瘤病毒(HPV)感染密切相关, 主要以HPV 16型感染为多见;另一类为HPV不相关的上皮内瘤变, 这类癌前病变的病理发生机制目前还在研究中。局部慢性非感染性炎症性外阴皮肤病, 如棘层肥厚性皮肤病的慢性单纯性苔藓和苔藓样皮肤病的硬化性苔藓等组织学表现可见于这类病变。根据p53表达的最新研究, HPV不相关的上皮内瘤变可分为:p53突变型, 即分化型外阴上皮内瘤变(differentiated-type vulvar squamous intraepithelial neoplasia, dVIN)和p53野生型, 即分化性外生型外阴上皮内病变和外阴棘皮病伴分化改变。以上所述上皮内瘤变或上皮内病变不仅在临床表现上不同, 早期病理变化细微, 在分子发生机制上也截然不同。因此, 此类病变在病理诊断和临床处理方面均具挑战。本文阐述并总结了目前dVIN的国内外病理研究进展和临床意义及临床靶向治疗的选择。     

女性下生殖道癌临床病理特征与人乳头状瘤病毒型别间的相关性研究

目的：探讨女性下生殖道癌的临床病理与人乳头状瘤病毒（HPV）型别之间的关系。方法：回顾性研究100例下生殖道癌（宫颈癌63例,外阴癌37例）的临床病理特征,并应用PCR技术检测每份标本的HPV状态。结果：在模板内参照阳性的87份中,宫颈癌54份,外阴癌33份,HPV阳性率在宫颈癌中为88．3％,以HPV16型（55．6％）和HPV18型（24．4％）为主,在33例外阴鳞癌中,HPV阳性仅见于基底细胞癌和混疣样癌,阳性率均为83．3％,以HPV16型为主（70．0％）;6例基底细胞样癌中有3例合并宫颈鳞状上皮肿瘤,其中2例宫颈与外阴的肿瘤均为HPV16型阳性;21例角化鳞癌则未检测出HPV－DNA,但有发病年龄高（平均63．3岁）。形态学上角化明显和预后较差等特征。结论：HPV16型和18型在宫颈癌中性阳率较高,而在外阴癌中HPV阳性的意义则因组织学类型而不相同。     

6例分化型外阴上皮内瘤变临床病理观察

目的：探讨分化型外阴上皮内瘤变( differentiated-type vulvar intraepithelial neoplasia, dVIN)的临床病理学特征、鉴别诊断、治疗及预后。方法回顾性分析6例dVIN的临床特点、病理形态学和免疫表型特征,并复习相关文献。结果患者均为女性,年龄53~80岁,平均62岁,临床特点大多表现为外阴白斑、激惹、瘙痒、疼痛、溃疡、触血。病理组织学特征表现：基底与副基底层显著增生,上皮角下延、吻合,细胞显著异型,核仁明显,核分裂活跃,伴异常角化;中、表层细胞高度分化,细胞间桥显著,伴明显的胞质嗜酸性改变,表层角化过度与角化不全;邻近上皮真皮水肿与胶原化,带状淋巴细胞浸润,表皮增生与角化过度。免疫表型：p53在dVIN中的阳性率为83．3%(5/6),p16不表达(0/6),dVIN基底层和副基底层细胞Ki-67增殖指数>90%。4例随访时间为6~36个月,平均17个月,其中1例术后9个月死亡,1例术后6个月复发,复发和死亡的2例同时或异时伴浸润性鳞状细胞癌,其余2例术后分别随访18个月和36个月,均无复发。结论 dVIN是一种少见的高级别外阴上皮内病变,具有高的、潜在进展危险性,p53、p16及Ki-67联合使用有助于dVIN的诊断。     

外阴硬化萎缩性苔癣治疗前后的光镜与电镜观察

<正> 作者对17例外阴硬化萎缩性苔癣及治疗后有显效者8例进行光、电镜观察,另检查了3例外阴正常者作为对照。治疗前硬萎可见上皮细胞萎缩,细胞间隙增大(基底细胞间隙增大明显),细胞表面微绒毛与细胞间隙标粒均减少,细胞内黑色素颗粒减少或消失,细胞核周围有空泡形成。个别基底细胞膜有轻度缺损。基底细胞伸向真皮的微突,     

Lectin histochemistry for detection of the oligosaccharides in the testis of the chick embryo

The oligosaccharidic content, distribution and changes of the glycoconjugates in the testis of the chick embryo, from the 8th day of incubation to hatching, were studied using a battery of seven HRP-conjugated lectins (DBA, SBA, PNA, ConA, WGA, LTA, and UEA I). Our findings showed that ConA and WGA appeared to characterize the spermatogonia during their differentiation, maturation, migration and meiosis. In the Sertoli cells, a change of localization of staining with ConA and WGA was revealed during the differentiation and maturation of these cells. The basal membrane was characterized by the reactivity with ConA and WGA from the early stages of incubation. ConA, WGA and PNA reacted with the endothelial cells of the testis for the whole period of incubation considered. Moreover, the interstitial cells, since their appearance, showed reactivity with ConA, WGA and PNA at the plasma membrane and the cytoplasm.     

Synovial sarcoma--a misnomer.

For an evaluation of the putative histogenetic relationship of synovia and synovial sarcomas, normal synovia, villonodular synovitis, and synovial sarcomas were compared for their patterns of expression of intermediate filaments of keratin and vimentin type and epithelial membrane antigen and for lectin binding sites. The lining cells in both normal synovia and villonodular synovitis reacted with anti-vimentin antibodies, but not with antibodies to different types of keratins or epithelial membrane antigen. The cleft-lining cells in synovial sarcoma, on the other hand, showed only keratin positivity, and epithelial membrane antigen could also be detected in these cells. Nonneoplastic synovial lining cells bound peanut agglutinin (PNA), Ricinus communis agglutinin (RCA), soybean agglutinin (SBA), concanavalin A (Con A), and wheat germ agglutinin (WGA) conjugates, but not Ulex europaeus I lectin (UEA I). In contrast, the epithelial-like cleft lining cells in synovial sarcomas showed an apical cytoplasmic binding of PNA, UEA I, RCA, and SBA, and binding of WGA to the whole cytoplasm but did not bind Con A. The distinct differences between synovial lining cells and synovial sarcoma cells speak against synovial cell features in synovial sarcoma. These results indicate that synovial sarcoma is a carcinosarcomalike tumor with true epithelial differentiation, and the term "synovial sarcoma" apparently is a misnomer that should be abandoned.     

A study of lectin bindings to the fetal membranes.

Human tissues contain carbohydrates for a main component, functioning as a source and reservoir of energy, connective and supporting element, recognition site and related tasks. Our main interest is to reveal the synthesis and distribution of carbohydrate elements in human fetal membranes. The aim of our work was to clarify, which kinds of elements containing carbohydrates, existed in the fetal membranes. Therefore we applied a lectin-binding study using the following FITC labelled lectins: ConA, WGA, PNA, LCA, RCA. This lead to the result, that ConA, LCA, WGA and RCA produced a positive reaction in the amnion epithelium, which was negative when using PNA. The basement membrane I showed an intense fluorescence when we used ConA, LCA and WGA, using RCA it was weaker and using PNA fluorescence was nearly missing. The examination of the amniotic fibroblast and intercellular substance showed a positive reaction with all lectins, but the intercellular substance lead to weaker fluorescence. The chorionic fibroblasts, intercellular substance and basement membrane II produced fluorescence using ConA, LCA, WGA and PNA, but no reaction could be examined, when using RCA. The trophoblastic cells did not react with LCA and RCA. The intercellular substance reacted positively with all lectins.     

Lectin and antibody labelling of developing rat photoreceptor cells: an electron microscope immunocytochemical study

Summary Lectin and rhodopsin antibody binding sites were studied in developing and adult rat photoreceptors in order to compare changes in the total carbohydrate pool with the movement of a known glycoprotein rhodopsin. Electron microscope immunocytochemical techniques utilizing modified colloidal gold methods were used. At birth, all three lectins-Concanavalin A (ConA), Ricinus communis agglutinin II (RCA II) and wheat germ agglutinin (WGA) — showed heavy labelling of the photoreceptor surface scierai to the outer limiting membrane. At the same age, a monoclonal antibody against rhodopsin, RET-P1, revealed sparse labelling of only occasional immature photoreceptor surfaces. At postnatal day 4(P4), all three lectins showed variable binding to the inner segment and along the length of the newly forming connecting cilium. There was generally a region of more intense label at the base of the cilium. RET-P1 binding to P4 retina showed a discontinuous distribution, with heavily labelled inner segments being adjacent to unlabelled inner segments. This pattern indicates that the initial expression of rhodopsin is not a coordinate event but occurs in discrete cells, possibly related to the end of mitosis. RET-P1 binding at this age was reduced or absent from the proximal connecting cilium. AT P7, when the outer segments are beginning to develop, all the lectins and RET-P1 showed reduced binding to the inner segment plasma membrane and heavy labelling of the outer segment surface. In favourable sections, heavy labelling of the photoreceptor cell body plasma membrane by ConA and RCA II was also observed, terminating abruptly at the outer limiting membrane. The variation in ligand binding between different cellular compartments which are all formed from a continuous plasma membrane may indicate the presence of special barriers to diffusion of membrane components. This labelling pattern persisted into maturity. RET-P1 and lectin binding did not always correspond in developing retina, indicating that at least part of the observed lectin label must be due to other glycoproteins or glycolipids. Post-embedding thin section labelling of adult rat retina revealed a uniform binding pattern across the outer segment for ConA, WGA and RET-P1. However, RCA II exhibited labelling only along the basal edge of outer segments. Labelling of isolated, opened discs from bovine rod outer segments revealed binding to a single surface for ConA, WGA and RET-P1, but RCA II only labelled a small amount of membrane. Hence RCA II seems to recognize a determinant present only on the outer segment plasma membrane.     

<Emphasis Type="Italic">Fasciola hepatica</Emphasis> miracidia: Lectin binding and stimulation of <Emphasis Type="Italic">in vitro</Emphasis> miracidium-to-sporocyst transformation

The lectin binding properties of Fasciola hepatica miracidia were studied by a panel of fluorescein- and gold-conjugated lectins (ConA, LCA, WGA, LEA, SBA, HPA and UEA-I). The presence of mannose and/or glucose residues was demonstrated with ConA and LCA as weak diffuse fluorescence of the miracidial surface, which was more intense at the anterior part of the larva. The N-acetylglucosamine-binding lectins WGA and LEA reacted intensely with the whole miracidial surface. No labelling with N-acetylgalactosamine and/or galactose-specific (SBA and HPA) and fucose-specific UEA-I lectins was observed. The possibility that the specific recognition of the miracidial surface carbohydrates by lectins may initiate the process of transformation of the miracidia into sporocysts was examined in vitro in physiological saline for Galba truncatula. Incubation in the presence of ConA and WGA resulted in facilitation of the transformation process. Facilitation was absent in the presence of inhibitor sugars. Incubation in the presence of SBA or UEA-I had no effect. The results suggested a possible impact of carbohydrate-lectin interactions in transformation of miracidia of F. hepatica to sporocysts in vivo.     

Lectin binding patterns in benign and malignant lesions of the breast.

The binding patterns of a panel of eight lectins were studied in twenty eight breast lesions, consisting of ten cases of fibroadenoma, three cases of cystosarcoma phyllodes, five cases of fibrocystic disease and ten cases of infiltrating duct carcinoma by light microscopy. The eight lectins viz. PNA, WGA, RCA, SBA, UEA I, LTA, LCA and Con A were tested on paraffin sections using the Avidin Biotin Peroxidase Complex technique. PNA, RCA and UEA I showed a consistent positivity in benign and malignant lesions. The binding was localised mainly along the apices of mammary ductal epithelial cells in the benign lesions. In contrast, the malignant cells showed a considerable variation in staining patterns like diffuse cytoplasmic, membranous, and vacuolar. No definite correlation was seen between the intensity of binding and the histological grade in infiltrating duct carcinoma except in the case of Con A which was seen to bind more intensely to poorly differentiated tumours. The diagnostic significance of these patterns have been discussed.     

Molecular identification of lectin binding sites differentiating related low and high metastatic murine lymphomas

We have previously shown that differences in cell surface carbohydrates can be detected in related murine tumor lines of varying metastatic capacity using plant lectins such as soybean agglutinin (SBA) orVicia villosa (VV) but not concanavalin A (ConA). Here we show that weakly metastatic Eb cells bind SBA via four glycoproteins and one GL2-like glycolipid. The major high-affinity SBA binding component of weakly metastatic ESb-MP cells was a glycoprotein of 210–220 kd. Highly metastatic ESb cells also expressed this protein but the oligosaccharide side chains were altered in such a way that SBA-binding was completely lost while ConA and peanut agglutinin (PNA) binding remained similar. Quantitative binding studies using iodinated lectins indicated that SBA binding of ESb cells could only be detected at lectin concentrations > 75g/ml. The role of altered carbohydrates in metastasis is discussed.     

The stimulation of human B and T lymphocytes by various lectins

The response of human peripheral blood lymphocytes to different lectins was tested in vitro by monitoring DNA synthesis, blast transformation, and mitotic activity. One group of lectins - RCA, VGA, HPA, PNA, and UEA - showed no stimulating effects at all. WGA and VVA induced DNA synthesis and blast transformation but failed in stimulating mitosis. The mitogens PHA, ConA, LCA, and PWM showed peaks of mitotic activity at 50-60 hours for PHA, 70 hours for ConA, 80 hours for LCA, and between days 4 and 5 for PWM. The stimulation of different subpopulations of lymphocytes was investigated by immunological methods for the detection of B- and T-cell-specific surface structures during the whole incubation period. PHA proved to be a predominantly T cell stimulating agent, whereas ConA seemed to activate a higher proportion of B cells than yet known. PWM and the so-called T cell mitogen LCA turned up to stimulate a large number of B cells, but lead also to a T cell activation. The analysis of SCE events in stimulation experiments with these two lectins showed the early proliferation of a cell population with low SCE frequencies and the late propagation of a cell population with higher SCE rates. It could be assumed that the first population is represented by B- and the second by T-cells.     

Lectin cytochemistry of carbohydrates on cell membranes of rat cerebellum

Summary Wheat germ agglutinin (WGA),Ricinis communis agglutinin (RCA) andLens culinaris (LC) lectins have been used to characterize carbohydrates of neuronal membrane systems in rat and chick cerebellum. WGA and RCA both label Golgi membrane cisternae on the side of the membrane facing the cisternal space but they do not label other elements of the granular and agranular endoplasmic reticulum (ER). WGA labels the plasma membrane generally and WGA binding sites are concentrated within the synaptic cleft and at specialized sites along the axolemma at the node of Ranvier. RCA labelling of plasma membranes is sparse. Neuraminidase treatment of tissue slices prior to lectin cytochemistry does not alter the Golgi membrane distribution of WGA and RCA binding sites and other ER membranes remain unlabelled. The plasma membrane staining with WGA is decreased and that with RCA is increased after neuraminidase pretreatment. The pattern of lectin labelling with LC resembles that seen with concanavalin A (con A) in that all elements of the ER within cell bodies label on the side of the membrane facing the cisternal space. A major difference when compared to con A labelling is that the hypolemmal cisternae lying adjacent to the plasma membrane of Purkinje cell axons and dendrites do not label with LC. Collectively, these results suggest that elements of the ER of Purkinje cells are heterogeneous with respect to their lectin binding properties in a manner which is consistent with the formation of more complex oligosaccharides on membranes of the cell periphery.     

Histochemical Analysis of Changes in Lectin Binding in Murine Glomerular Lesions

Lectin binding in diseased murine glomeruli was studied in MRL 1 mice, using seven different fluorescence- or peroxidase coupled lectins: Griffonia simplicifolia I (GS-I) , Ulex europaeus agglutinin I (UEA-I) , Ricinus communis agglutinin I (RCA I), wheat germ agglutinin (WGA), concanavalin A (Con A), peanut agglutinin (PNA), and Helix pomatia agglutinin (HPA). Lectin binding in diseased glomeruli of MRL 1 mice was different from that in normal glomeruli. Light and fluorescence microscopy showed that: 1. in mesangial proliferative lesions, the binding of RCA I, WGA and Con A increased and that of GS I and PNA appeared in the mesangium; 2. in other glomerular lesions, UEA-I bindng appeared and RCA I stained the altered membranes irregularly. Electon microscopy showed that: 1. GS-I stained the endothelial cell coat and the glomerular basement membrane covered by the endothelial cells; 2. GS I strongly stained the dilated subendothelium in regions of mild mesangial interposition; 3. GS-I stained the cell coat of invasive macrophages; 4. GS-I and UEA-I stained the cell membrane-like material derived from degenerative endothelial cells; 5. RCA I stained the epithelial and endothelial cell coats and the glomerular basement membrane. These results indicate that lectin-binding studies can be used for analysis of glomerular lesions.