涎腺粘液表皮样癌凝集素检测意义

本文应用组织化学和免疫组织化学方法,研究6例粘液表皮样癌,9例正常涎腺作对照观察。用4种凝集素抗体(PNA、WGA、SBA 和 UEA)检测正常细胞和肿瘤细胞,结果表明,PNA 和 UEA 在细胞分布上极不一致,用于诊断涎腺粘液表皮样癌有实用意义,而癌内粘液属混合性粘蛋白,与正常腺体内粘液相比,仅含量略增高。凝集素检测本癌还可解释癌细胞发生、转化的部分原因。     

凝集素受体在胃癌及其淋巴结转移癌中的分布

目的：探讨凝集素受体分布与胃癌及其淋巴结转移癌的关系。方法：应用生物素标记的3种凝集素（Biotin-PHA,Biotin-PNA,Biotin-DBA)对人体胃癌66例,淋巴结转移癌32例,正常胃粘膜10例,进行亲合组织化学法（ABC法）研究。结果：发现PHA、PNA受体的分布与胃癌组织学类型及分化程度有关。PHA、PNA在胃癌中标记阳性率较高,对于胃癌的诊断是一个十分有用的探针。81．3％淋巴结转移癌与其原发胃癌在凝集素标记的量上存在着差异,并且比原发肿瘤获得更多的PHA受体。PHA、PNA在淋巴结转移癌中的标记阳性率较高,对于识别淋巴结是否转移具有一定的应用价值。结果：胃癌中凝集素受体分布与其组织学类型及淋巴结转移癌有关。     

癌胚抗原和凝集素受体在胆囊病变中的观察

用癌胚抗原（CEA）和4种凝集素（RCA、UEA、PNA、PHA）对48例睑囊切除标本进行免疫组织化学观察，发现CEA在胆囊的正常、增生、异常增生和癌上皮中阳性率和阳性强度呈递增趋势。4种凝集素受体则在胆囊化生上皮中呈不同程度阳性反应。结果表明，CEA可作为检测胆囊癌的较敏感的标志物，4种凝集素对识别癌前病变和胆囊癌功能分型有一定参考价值。     

大肠息肉粘液组织化学和免疫组织化学研究

应用 HID/AB 组织化学染色法,CEA 免疫组化及凝集素亲合组织化学方法对56例大肠息肉研究发现:41例肿瘤性息肉(简称腺瘤)中,HID/AB 粘蛋白染色3例出现唾液酸粘蛋白,且为中、重度不典型增生,而3例化生性息肉有2例显示唾液酸粘蛋白着色。伴有轻、中、重度不典型增生的腺瘤 CEA 阳性率分别为75%、84.6%和100%。WGA、DBA、UEA 和 PNA4种凝集素染色在腺瘤和非肿瘤性息肉间无明显差别。但轻、中和重度不典型增生的腺瘤 PNA 阳性率则分别高达60%、69.2%和100%。化生性息肉3例 PNA 染色均阳性。因此认为唾液酸粘蛋白、CEA 和与 PNA 结合的糖蛋白可能为癌变高发群的生物学标记,且化生性息肉与癌发生的关系值得重视。     

检测胆汁糖蛋白糖链结构变化鉴别良恶性胆道疾病分析

目的：探讨胆汁糖蛋白糖链结构变化对于鉴别良恶性胆道疾病的作用。方法选取青岛市海慈医疗集团普外科收治的胆道疾病患者100例,按照良、恶性将其分为对照组（良性胆道疾病）和观察组（胆管癌）,每组各50例。取两组患者胆汁滴于硝酸纤维膜上,通过比较麦胚凝集素（ WGA ）、欧曼陀罗凝集素（ DSA）、小扁豆凝集素（ LCA）、刀豆凝集素（ CONA）试验阳性率,探讨胆汁糖蛋白糖链结构变化与良恶性胆道疾病的关系。结果对照组 WGA、DSA、LCA、CONA 凝集素试验阳性率分别为22．0％、14．0％、2．0％、76.0％,观察组分别为76．0％、66．0％、76．0％、82．0％。两组CONA凝集素试验阳性率差异无统计学意义（P＞0．05）,观察组WGA、LCA、DSA凝集试验阳性率均明显高于对照组,差异均有统计学意义（χ2＝29．17、28．17、57．55,均P＜0．05）。结论胆汁糖蛋白糖链结构变化与胆道疾病良、恶性密切相关,可以通过WGA、LCA、DSA凝集试验阳性率判断胆管疾病良、恶性,值得在临床上广泛推广。     

凝集素在食管上段胃黏膜异位症中的表达及意义

目的比较不同种类凝集素在食管上段异位胃黏膜及正常食管组织中表达,利用凝集素受体在食管黏膜分布情况来推断食管上段异位胃黏膜的来源。方法收集取自2011年1月至2015年12月山西医科大学汾阳医院消化内科47例食管上段胃黏膜异位症患者组织及其相应的正常食管组织;分别以DBA,LCA,BSA一抗,应用链霉卵白素过氧化物酶法(SP法)进行免疫组织化学染色;观察DBA、LCA、BSA在食管上段异位胃黏膜组织中的表达特征。分析食管上段胃黏膜异位症患者临床特征。结果与正常食管组织及胃窦组织比较,DBA、LCA、BSA 3种凝集素表达减少。与食管下段Barrett食管组织比较,3种凝集素表达下降。结论 DBA、LCA、BSA 3种凝集素可能在食管上段异位胃黏膜组织发生、发展中起作用,为进一步研究食管上段胃黏膜异位症的发生机制提供思路。     

12例鼻息肉中花生凝集素受体的研究

近 2 0年来凝集素在生物学方面的应用研究发展尤为迅速 ,它最大的特点在于其能够识别糖和糖类 ,每一种凝集素具有对某一种特殊的碳水化合物有专一结合的能力 [1 ] ,凝集素受体是指能与相应凝集素特异结合的糖复合物 (糖蛋白 ,糖脂和蛋白聚糖 ) ,它在细胞的多种功能中起主要作用 ,如细胞间的识别和粘着、营养吸收、抗原抗体反应等 [2 ] 。本文利用花生凝集素 (PNA)与其糖复合物受体特异结合的特性 ,采用免疫组化ABC技术研究鼻息肉中花生凝集素受体的分布及其意义。1　材料与方法1.1　材料 :经病理证实的鼻息肉标本 12例 ,正常鼻窦粘膜标本…     

荷人脑瘤裸小鼠模型（NHG—1）的移植瘤染色体分析

为探讨荷人脑恶性胶质瘤裸小鼠模型长期体内生长传代过程中移植瘤的染色体特性，我们对SHG－44第67代细胞及由该细胞接种于裸小鼠以后建立的NHG－1第1、22、35、40、45和50代移植瘤细胞进行染色体分析。结果染色体分布，SHG－44第67代细胞超三倍体染色体高达83％；NHG-1第1代移植瘤细胞仍以超三倍体染色体为主，占46％；第22代移植瘤细胞则以亚三倍体染色体为主，占70％；第35代以后各代移植瘤细胞均以亚二倍体染色体为主，染色体分布稳定。染色体核型，SHG－44细胞10个G显带分析，均可见到3条较长的亚中部着丝点标记染色体。NHG－1移植癌细胞每代5个细胞G显带核型分析，发现存在着人脑染色体。     

ABO血型不合的骨髓移植

HLA相配的骨髓移植（BMT）是治愈白血病、再障、免疫缺陷病等有效甚至唯一的疗法。ABO血型抗原不依赖HLA基因复合体遗传[1]，故25％HLA相配的同胞约15～20％供受体ABO血型主要不合[2]。血型不合与BMT的植活、排斥、急性移植物抗宿主病（GVHD）发生率及严重度、生存时间无关，而其主要问题是：移植时预防溶血和移植后输血选择。ABO血型不合分两类：（1）主要不合：受者有与供者红细胞抗原起反应的红细胞凝集素。（2）次要不合：供者有与受老红细胞抗原起反应的红细胞凝集素。一、预防溶血次要不合的BMT可按常规进行。如供者血球…     

米非司酮对乳腺癌细胞系MCF-7/ADM裸鼠移植瘤耐药逆转作用

目的：探讨米非司酮对乳腺癌细胞系MCF-7/ADM裸鼠移植瘤耐药逆转作用。方法：以人乳腺癌细胞系MCF-7/ADM建立耐阿霉素人乳腺癌裸鼠皮下移植瘤动物模型,将成瘤裸鼠随机分为空白对照组,米非司酮组（MIF组）,阿霉素组（ADM组）,米非司酮联合阿霉素组（MIF＋ADM组）。测量裸鼠移植瘤横径与短径,计算移植瘤体积,绘制生长曲线。结果：对裸鼠干预处理4周后,MIF＋ADM组移植瘤体积[（232.5149±309.2377）mm3]最低,ADM组[（508.9648±16.2609）mm3]次之,均低于空白对照组移植瘤体积[（962.2309±261.1313）mm3],差异有统计学意义（P〈0.05）;MIF＋ADM组移植瘤体积较单独用药的MIF组及ADM组低,差异具有统计学意义（P〈0.05）。结论：米非司酮有逆转乳腺癌细胞MCF-7/ADM裸鼠移植瘤耐药性的作用。     

Antitumor effects of concanavalin A and Sophora flavescens lectin in vitro and in vivo

Aim： Proteins with legume lectin domains are known to possess a wide range of biological functions. Here, the antitumor effects of two representative legume lectins, concanavalin A （ConA） and Sophora flavescens lectin （SFL）, on human breast carcinoma cells were investigated in vitro and in vivo. Methods： Human breast carcinoma MCF-7 cells and human normal mammary epithelial MCF-IOA cells were examined. Cell viability was detected using WST-1 and CCK-8 assays. Cell apoptosis was analyzed with Hoechst 33258 staining. Cell cycle was investigated using flow cytometry. The expression of relevant proteins was measured using Western blotting. Breast carcinoma MCF-7 bearing nude mice were used to study the antitumor effects in vivo. The mice were injected with ConA （40 mg/kg, ip） and SFL （55 mg/kg, ip） daily for 14 d. Results： ConA and SFL inhibited the growth of MCF-7 cells in dose- and time-dependent manners （ICso values were 15 and 20 pg/mL, respectively）. Both ConA and SFL induced apoptotic morphology in MCF-7 cells without affecting MCF-IOA cells. ConA and SFL dose- dependently increased the sub-G1 proportion in MCF-7 cells, while SFL also trip=~ered the G2/M phase cell cycle arrest. Both ConA and SFL dose-dependently increased the activities of caspase-3 and caspase-9 and release of cytochrome c from mitochondria into cytoplasm, up-regulated Bax and Bid, and down-regulated Bcl-2 and BcI-XL in MCF-7 cells. ConA reduced NF-KB, ERK, and JNK levels, and increased p53 and p21 levels, while SFL caused similar changes in NF-KB, ERK, p53, and p21 levels, but did not affect JNK expression. Administration of ConA and SFL significantly decreased the subcutaneous tumor mass volume and weight in MCF-7 bearing nude mice. Conclusion： ConA and SFL exert anti-tumor actions against human breast carcinoma MCF-7 cells both in vitro and in vivo.     

Plant lectins are novel Toll-like receptor agonists

The T cell mitogen and plant glycoprotein, phytohaemagglutinin (PHA), is commonly used to stimulate peripheral blood mononuclear cell (PBMC) preparations to produce IL-2, IL-5, GM-CSF and IFN-γ and so provide an assay to detect immunosuppressants like FK506 and anti-inflammatories such as PDE IV inhibitors.During the early discovery of novel TLR agonists for the treatment of asthma we initially showed that PHA-L is a specific human TLR4 agonist, devoid of effects on equivalent TLR4 null cells. This TLR4 agonism was not due to LPS contamination of PHA-L, as polymyxin B was ineffective and unlike PHA-L, LPS did not stimulate TLR5 or TLR2/6. Also this specific PHA-L agonism of TLR4 was shown for different PHA forms, for example, PHA-P.This TLR lectin pharmacology finding was further explored by testing a broader panel of plant lectin representatives for agonism against a suite of hrTLR cell reporter assays (2/6, 3, 4, 5, 7, 8 and 9).Soybean agglutinin (SBA), concanavalin A (ConA) and PHA lectin family members only stimulated extracellular TLRs (2/6, 4 and 5) probably due to lack of intracellular access, whilst other lectins were either pan-active (WGA) or inactive (AIL).Interestingly SBA only stimulated TLR4, ConA, TLR2/6 and PHA-L, TLR2/6, 4 and 5. As each lectin family exhibits different sugar ligand specificity for interaction, these results suggest that the pharmacology of this TLR agonism is encoded by the lectin's carbohydrate recognition motifs and the appropriate surface presentation of these motifs on different TLRs.     

Antiproliferative effects of lectins from Canavalia ensiformis and Canavalia brasiliensis in human leukemia cell lines

The antiproliferative activity of lectins Canavalia ensiformis (ConA) and Canavalia brasiliensis (ConBr) were studied using human leukemia MOLT-4 and HL-60 cell lines. It was revealed that both ConA and ConBr were markedly cytotoxic to cells using MTT and NAC assays. The IC₅₀ values were approximately 3 and 20 μg/mL for ConA and ConBr, respectively, for both MOLT-4 and HL-60 cells. However, in normal human peripheral blood lymphocytes, the lectins were not cytotoxic, even when tested at concentrations as high as 200 μg/ml. Using comet assay, the lectins produced a rate of DNA damage exceeding 80% in MOLT-4 and HL-60 cells. Fluorescence analysis revealed the morphology characteristic of apoptosis, with low concentrations of apoptotic bodies and fragmented DNA (5 μg/ml). Flow cytometric analysis demonstrated an accumulation of cells in the sub-G1 cell cycle that is characteristic of DNA fragmentation, and a decrease in membrane integrity at high concentrations. Lastly, we evaluated the alterations in mitochondrial potential that reduced after treatment with lectins. Our results indicate that ConA and ConBr inhibited cell proliferation selectively in tumor cells and that apoptosis was the main death mechanism. Therefore, lectins can be considered a class of molecules with a high antitumor activity potential.     

原代人结肠癌裸鼠皮下移植瘤模型的建立

目的建立人结肠癌裸鼠皮下移植瘤模型。方法采用组织块直接贴壁法将新鲜人结肠癌组织进行原代细胞培养,反复贴壁法和胰酶消化法使之纯化。纯化后的结肠癌细胞接种于裸鼠左侧腋处皮下。观察成瘤时间、肿瘤生长情况、移植肿瘤大小及裸鼠摄食、活动状况。实验结束后取移植瘤以HE染色和CEA及CK20免疫组化检验。结果本实验原代结肠癌细胞可以在裸鼠皮下存活,接种后第8天见有移植瘤形成,成瘤率为80%。病理切片显示组织细胞排列成不规则腺体状,有核分裂相。CEA及CK20检验阳性。结论初步建立了人结肠癌的裸鼠皮下移植瘤模型,为进一步研究结肠癌奠定了基础。     

Light and electron microscopy investigation of glycoconjugates of rat hippocampus by lectin-gold technique

The distribution of glycoconjugates in rat hippocampus was investigated by light and electron microscopy using gold-labelled lectins with different sugar specificities: ConA, WGA, PNA and LTA. A morphological correlation with cytochemical data was performed, and revealed the presence of intensely-stained (dark), unstained (light) and weakly-stained (intermediate) neurons both in Ammon's horn and in the dentate gyrus. The glycoconjugates are located in the nucleus, the perikaryal cytoplasm and the cytoskeleton of axons and dendrites: they are probably involved in the mechanisms of transport towards the synaptic membrane. The presence or absence of glycoconjugates corresponds to the particular ultrastructural aspect of the cell. This suggests the existence of different functional states of the neurons, probably correlated with the synthesis of neurotransmitter precursors or receptors.     

Discrepancy between in vitro and in vivo passaged U-937 human leukemic cells: tumorigenicity and sensitivity to differentiating drugs

The treatment of nude mice bearing tumors of transplanted human leukemic cells with drugs known to induce differentiation of the same leukemic cells in vitro does not always affect tumor yield, tumor cell differentiation or nude mice survival. We have transplanted human monoblastic leukemic cells of the U-937 cell line into newborn Swiss nu/nu mice. Priming with cyclophosphamide, followed by subcutaneous injections of at least 10 x 10(6) cells allowed us to obtain solid tumors. The cytology, HLA phenotype and in vitro proliferation characteristics of the U-937 tumor cells were conserved. However, these tumor cells were more tumorigenic when reinjected into nude mice and showed a modified response to differentiation induction. A decreased capacity to differentiate with retinoic acid (RA) and a resistance to 1-beta-D arabinofuranosyl cytosine (Ara-C) and 1-25 dihydroxy vitamin D3 (1-25 (OH)2 D3) were noted in three tumor cell lines tested. With regard to the latter, the resistance was not due to a modification of the number of cell receptors. The study shows that though in vivo transplantation of human leukemic cells in nude mice may lead to a selection of resistant cells, systematic checking of in vitro differentiation characteristics of the tumor cells permits the nude mouse model to be maintained for the in vivo screening of new differentiating agents.     

Binding characteristics of 3H-dihydroalprenolol to beta-adrenergic receptors of rat brain: influence of lectins

The significance of the carbohydrate moieties of the beta-adrenergic receptor molecule in the rat brain was examined using the radioligand binding assay method. Thus, this experiment was designed to assess the effects of lectins, concanavalin A (Con A), Phaseolus vulgaris agglutinin (PHA), and wheat germ agglutinin (WGA) on the affinity of the beta-adrenoceptor. The rat brain was used and the beta-adrenoceptor binding assay was carried out using 3H-dihydroalprenolol as a ligand. Con A and PHA significantly caused an increase in the values of the density of beta-adrenoceptor (Bmax) and a reduction in the values of the dissociation constant (Kd), but significant changes were not observed with WGA. These results strongly suggest that the carbohydrate moieties of the cell surface containing the beta-adrenoceptor molecule may have a crucial role in the drug-receptor interaction, and they imply that the beta-adrenoceptor molecule is a glycoprotein which contains N-linked carbohydrate chains.     

个体化裸鼠荷人肺癌肿瘤模型的建立

目的:建立人肺癌个体化可传代组织块动物模型.方法:组织块移植组将直径2mm的人肺癌新鲜标本癌组织块移植于BALB/C裸鼠背部皮下组织内,每例人肺癌新鲜标本均移植入5只裸鼠体内,成瘤者为第1代.取第1代移植瘤组织块以相同方法每例标本移植于另5只BALB/C裸鼠背部皮下组织内,成功者为第2代.重复上述传代方法得到第3代移植瘤.将人肺癌细胞株A549、H1795分别移植于BACLC裸鼠背部皮下组织内,成瘤达500 mm3后取癌组织块.分别取实验组的原代人肺癌肿瘤及第1、2、3代移植瘤部分组织,及作为对照的细胞系移植瘤组织,固定于10％福尔马林溶液中,石蜡包埋、切片、HE染色,光镜下观察比较其形态异同.结果:组织块移植组瘤细胞不仅保留了人肺癌细胞的异型性,还保持原有排列结构；而作为对照的细胞系移植瘤细胞失去了原有的排列结构,仅保留了癌细胞的异型性.结论:利用不同患者的手术切除肺癌标本所建立的裸鼠荷人肺癌肿瘤模型具有个体化特征,为肺癌个体化治疗的研究提供依据.     

人白细胞介素-24治疗人宫颈癌荷瘤裸鼠的实验研究

目的探讨人白细胞介素(hIL)-24基因重组质粒对人宫颈癌Siha细胞裸鼠移植瘤生长的抑制作用及其作用机制。方法建立荷宫颈癌Siha细胞裸鼠移植瘤模型,15只裸鼠随机分为3组:白细胞介素(IL)-24重组质粒组,空质粒组,磷酸盐缓冲液(PBS)组,根据肿瘤体积的生长情况绘制肿瘤生长曲线;处死裸鼠后切取瘤体并称质量,计算肿瘤抑制率;用流式细胞仪检测细胞周期。结果体内实验表明,IL-24重组质粒能够抑制肿瘤的生长,IL-24重组质粒组移植瘤的体积明显小于空质粒组和PBS组(P<0.05),空质粒组和PBS组移植瘤体积则差异无统计学意义(P>0.05)。IL-24重组质粒组抑瘤效果显著,其抑瘤率为56.5%,差异有统计学意义(P<0.05),空质粒组抑瘤率为1.3%,差异无统计学意义(P>0.05)。经流式细胞术检测,与空质粒组和PBS组比较,IL-24重组质粒组G0/G1期细胞增多(P<0.05),S期细胞减少,G2/M期细胞减少(P<0.05),细胞被阻滞在G0/G1期,空质粒组和PBS组则差异无统计学意义(P>0.05)。结论于荷瘤裸鼠的瘤内注射IL-24重组质粒可抑制肿瘤生长,提示IL-24具有明显的体内抗肿瘤作用。     

Distinct glycosylation in interstitial and serum tenascin-X

We developed an easy and fast method to isolate extracellular matrix tenascin-X (TNX) from various tissues in mice based on TNX antibody affinity purification. We purified approximately 350-kDa cellular interstitial TNX (iTNX) from the spleen, liver and kidney as well as 200-kDa serum TNX (sTNX). Since the nature and significance of glycosylation in TNX remains to be elucidated, glycobiochemical properties of purified TNX were characterized by lectin blot analysis. Lectin blots by Con A, LCA, PHA-E4, RCA120 or WGA revealed the presence of N-glycan in the cellular TNX and especially complex-type N-glycan in the serum TNX. In addition, the iTNX from liver and kidney also possessed O-glycan based on the reaction to PNA. The binding to AAL indicated that iTNX from the three tissues possesses fucose linked alpha1,6 to a pentasaccharide core, whereas sTNX does not. The reaction to SSA but not to MAM suggested the presence of sialic acid linked alpha2,6 to galactose in both cellular and serum TNX. Lectin blots of trypsin-treated iTNX from the spleen also demonstrated that WGA alone reacts to the t300 product derived from the amino-terminal 300-kDa portion.