先天性巨结肠肠壁神经组织免疫组织化学分析与检验学研究

目的研究先天性巨结肠肠壁神经组织免疫组织化学分析的诊断方法。方法设对照组,采用一组单抗及多抗试剂,对肠壁神经组织进行免疫酶标组织化学染色,光镜下观察其形态特征。结果①NSE染色:正常组阳性、HD病例狭窄段阴性;②S-100蛋白染色:正常组与HD病例神经细胞核染色阴性,而在巨结肠大量增生的神经丛染色阳性。结论神经元特异性烯醇酶(NSE)存在于神经元及轴突,S-100蛋白分布于神经纤维的雪旺氏细胞,二者形成清晰对比,可完全明确判断是否有神经元的存在和神经纤维增生的程度。     

胃肠道间质瘤病理及免疫组化分析

目的 探讨胃肠道间质瘤(GIST)的临床病理、免疫组织化学特点.方法 应用常规病理观察36例GIST的形态特征,用免疫组织化学SP法检测CD34、CD117、vimentin、SMA、NSE、S-100蛋白在GIST中表达.结果 本组GIST镜下瘤细胞以梭形细胞和上皮样细胞为主,或两种混合存在,成束状、栅栏状、旋涡状或巢状排列.36例中良性12例,交界性14例,恶性10例.免疫组织化学表型CD34和CD117阳性的阳性率分别为80.5%和94.4%,而vimentin、SMA、S-100蛋白在肿瘤向平滑肌或神经方向分化时阳性.结论 GIST由梭形细胞或上皮样细胞构成,组织结构形态多变,但免疫表型完全一致;CD117及CD34阳性可作为GIST诊断及鉴别诊断的重要参考依据.     

先天性肥厚性幽门狭窄S-100蛋白的免疫组化研究

目的通过对幽门部神经支持细胞特异性标记物S-100蛋白（S-100）的研究,进一步探讨先天性肥厚性幽门狭窄（congenital hypertrophic pyloric stenosis,CHPS）的发病原因。方法应用免疫组织化学染色观察48例CHPS患儿及10例正常对照组儿童幽门部神经支持细胞S-100蛋白表达。结果在正常对照组与CHPS环形肌及纵行肌中神经支持细胞S-100蛋白表达有显著差异,对照组为强阳性,实验组为阴性或弱阳性。结论神经支持细胞异常可能与幽门肥厚狭窄形成存在一定的关系。     

卵巢原发性纯类癌

报告1例卵巢原发性纯类癌。类癌细胞呈嗜银和亲银强阳性。免疫组织化学证实细胞内含有神经特异性烯醇化酶、突触蛋白、铬颗粒蛋白，Leu-7，5-羟色胺，人绒毛膜促性腺激素，细胞角蛋白，上皮膜抗原，癌胚抗原等物质，显示其特有神经内分泌，上皮性和癌胚性的多方向分化。     

肺硬化性血管瘤的临床病理特点

目的探讨肺硬化性血管瘤（SHL）的临床病理特点及组织起源。方法对15例SHL进行组织学观察及11种免疫组化标记：甲状腺转录因子-1（TTF-1）、上皮膜抗原（EMA）、广谱细胞角蛋白（CK／pan）、波形蛋白（Vimentin）、嗜铬素（CgA）、突触素（Syn）、神经元特异性烯醇化酶（NSE）、S-100蛋白、内皮细胞（Ⅷ因子、CD34）、间皮细胞（MC）。结果SHL主要由表面立方细胞和圆形、多角形细胞二种细胞类型组成；组织结构多样，主要表现为实性区、乳头区、血管瘤样区和硬化区四种结构。免疫组化标记二种细胞共同表达TTF-1和EMA；表面立方细胞表达CK／pan，不表达Vimentin；圆形、多边形细胞表达Vimentin，不达表CK／pan；CgA，Syn，NSE，S-100在少许圆形、多角形细胞中呈弱阳性表达；二种细胞均不表达Ⅷ因子、CD34及MC。结论SHL可能起源子原始未分化的呼吸上皮，可伴有神经内分泌细胞分化。     

食管鳞癌中Langerhans细胞（LC）的形态学观察

目的探讨S-100蛋白阳性LC形态的变化与食管鳞癌的发生发展及其分化程度、浸润深度和淋巴转移的关系。方法应用免疫组织化学SP法检测39例正常食管上皮、23例不典型增生及39例食管鳞癌中S-100蛋白的表达,观察不同食管黏膜组织中LC形态的变化。结果S-100蛋白定位于细胞浆。正常食管上皮主要表现为Ⅰ型LC,癌组织中主要表现为Ⅱ型LC;随着癌细胞分化程度的降低Ⅱ型LC逐渐增多;有淋巴转移者的Ⅱ型LC多于无淋巴转移者;在不同浸润深度的食管鳞癌中LC的形态无明显区别。结论LC形态变化与食管鳞癌的发生发展及其分化程度、淋巴转移有关,而与浸润深度无明显关系。     

食管鳞癌中Langerhns细胞(LC)的形态学观察

目的 探讨S-100蛋白阳性LC形态的变化与食管鳞癌的发生发展及其分化程度、浸润深度和淋巴转移的关系.方法 应用免疫组织化学SP法检测39例正常食管上皮、23例不典型增生及39例食管鳞癌中S-100蛋白的表达,观察不同食管黏膜组织中LC形态的变化.结果 S-100蛋白定位于细胞浆.正常食管上皮主要表现为Ⅰ型LC,癌组织中主要表现为Ⅱ型LC;随着癌细胞分化程度的降低Ⅱ型LC逐渐增多;有淋巴转移者的Ⅱ型LC多于无淋巴转移者;在不同浸润深度的食管鳞癌中LC的形态无明显区别.结论 LC形态变化与食管鳞癌的发生发展及其分化程度、淋巴转移有关,而与浸润深度无明显关系.     

视网膜母细胞瘤组织发生的探讨

应用免疫组织化学方法探讨视网膜母细胞瘤的组织来源。结果显示;神经胶质细胞标记物GFAP(12/25)、神经元标记物NSE(18/25)、S-100蛋白(10/25)瘤细胞呈不同程度的阳性反应。Vimentin瘤细胞全部呈阴性反应。结果提示:视网膜母细胞瘤来源于神经外胚层上皮,向神经元和光感受器方向分化。神经胶质细胞可能是反应性增生,但也不排除向神经胶质细胞方向分化的可能性。     

胃癌伴神经内分泌分化和胃混合性腺神经内分泌癌临床病理及预后分析

　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　　【摘要】目的 探讨胃癌伴神经内分泌分化和胃混合性腺神经内分泌癌的临床病理及预后。方法 回顾性分析61例胃癌伴神经内分泌分化和34例胃混合性腺神经内分泌癌患者的临床病理资料,对其组织化学及免疫组织化学染色进行观察。结果 胃癌伴神经内分泌分化和胃混合性腺神经内分泌癌患者肿瘤发病部位、远处转移及区域淋巴结转移差异有统计学意义（P<0.05）;SyN 、CgA和CD56三者阳性表达率差异有统计学意义（P<0.05）;区域淋巴结转移和远处转移2个因素与预后相关（P<0.05）;胃混合性腺神经内分泌癌患者术后生存期短于胃癌伴神经内分泌分化患者（P<0.05）。结论 免疫组化染色对胃癌伴神经内分泌分化和胃混合性腺神经内分泌癌的确诊具有重要意义,肿瘤神经内分泌细胞数量的多少对患者预后评估及辅助治疗有指导意义。      

免疫组织化学方法诊断先天性巨结肠症

目的 探讨免疫组织化学检查在诊断先天性巨结肠症中的价值。方法 应用神经细胞特异性烯醇化酶(NSE)和S_(100)蛋白对61例直肠粘膜吸引活检标本进行回顾性免疫组织化学检测。结果神经丛中NSE胞浆阳性深染的细胞和S_(100)蛋白染色的细胞状“空白”区可特征性地提示神经节细胞的存在,在巨结肠患者标本中则缺乏此类表现。33例临床诊断为巨结肠者,其手术切除标本经全层病理学检查均证实为巨结肠,诊断符合率为100％。结论 在直肠粘膜活检标本常规HE染色下诊断有疑问时,NSE结合S_(100)蛋白的免疫组织化学染色有助于确诊。     

Reversible rat mesenteric mast cell swelling caused by vagal stimulation or sham-feeding.

Fragments of rat mesentery were examined using acetylthiocholine to detect cholinergic nerve fibers and toluidine blue to identify mast cells. 59.2 +/- 2.6 percent of mast cells were at less than one-half mean cell diameter (4-5 microns), from the nerve fibers. Under the electron microscope, the membrane of mast cells was within less than 50 nm from axon membranes, suggesting a synaptic type of connection. Average mast cell area in fasted rats increased following feeding, stimulation of the left abdominal vagus nerve or exposure of the animal to the smell of food. It returned to control values within 60-80 min. Granule exocytosis was not observed. Mast cell swelling was prevented by atropine and induced by intravenously administered carbamylcholine. It appears that in rat mesentery, impulses travelling via cholinergic, parasympathetic fibers innervating mast cells, cause mast cell swelling. Compound 48/80 administered to rats at doses causing little degranulation and minimum release of histamine, caused extensive, reversible swelling of mesentery mast cells.     

Monoclonal murine anti-DNP IgE: in vitro histamine release of rat mast cells in the presence of reaginic rat and mouse sera

Mast cells from normal rats and animals reinfected with Nippostrongylus brasiliensis (N.b.) were sensitized in vitro with monoclonal anti-DNP mouse IgE, reaginic mouse serum with ovalbumin (OA) specificity and reaginic rat serum against N.b. The sensitized cells were triggered for the release of histamine with DNP-bovine-serum-albumin (DNP-BSA), OA, N.b.-homogenate and guinea-pig anti-mouse IgE. The histamine release from normal mast cells sensitized with monoclonal mouse IgE was inhibited either with N.b.-reaginic rat serum or OA-reaginic mouse IgE (30 micrograms/10(5) mast cells) completely desensitized mast cells from reinfected rats for specific histamine release in the presence of either N.b.-antigen or DNP-BSA. Anti-mouse IgE which was prepared by monoclonal mouse IgE did not bind to rat mast cells sensitized with rat IgE as was revealed by immunofluorescence experiments. Consequently we observed that anti-mouse IgE failed to trigger the histamine release from mast cells of reinfected rats.     

Mast cell subtypes from human lung tissue: their identification, separation, and functional characteristics

The contribution of mast cell subtypes and their different mediators to the pathogenesis of chronic obstructive lung diseases (COLD) has not yet been established. In the present study, enzymatic digestion, centrifugal elutriation and Percoll gradient centrifugation were used to obtain two populations of mast cell subtypes from human lung tissue. Mast cell subtypes were challenged with anti-human IgE, propranolol, compound 48/80, or opsonized zymosan. Both subtypes were able to release histamine, but differed in the amount of the amine release. Only the formalin-sensitive and alcian blue-positive type (FS-AB) released histamine on challenge with opsonized zymosan. The same subtype was able to release leukotriene C4 (LTC4) after challenge with anti-human IgE. The other subtype, the formalin-insensitive and alcian blue-positive type (FI-AB), did not respond to opsonized zymosan and did not release LTC4 after challenge with anti-human IgE. Stimulation with propranolol or compound 48/80 did not release histamine from the FS-AB mast cells while the FI-AB mast cells released only about 10% of their histamine content upon challenge with these secretagogues.     

克隆病P物质阳性神经组织及肥大细胞的形态学观察

目的：观察克隆病时Ｐ物质阳性神经组织、肥大细胞形态变化,并探讨其在该病发病中的作用。方法;应用免疫组织化学,组织化学方法对２７例克隆病（回肠１４例,结肠１３例）,１０例对照组肠壁Ｐ物质（ＳＰ）阳性神经纤维,神经元及肥大细胞进行观察。用形态计量学方法计量ＳＰ阳性神经组织的体密度及肥大细胞密度。结果：克隆病时肠壁粘膜下层神经纤维增生,部分呈“神经瘤样”改变,ＳＰ阳性神经元数量增多。粘膜下层及肌间神经丛     

Resveratrol inhibits the release of mediators from bone marrow-derived mouse mast cells in vitro

Resveratrol is a natural phytoalexin occurring in grapes, vines and peanuts and possesses both antitumor and antioxidation capabilities. Its chemoprotective actions are attributed partially to its anti-inflammation effect. The present study is aimed at checking the inhibitory actions of resveratrol on the release of mediators from bone marrow-derived mouse mast cells (BMMC) in vitro. Mast cells were prepared by isolating bone marrow cells from intact mice femora and culturing them for 4 weeks in the presence of IL-3 and FCS. The release reaction was triggered by IgE or calcium ionophore (A23187). Mediated by IgE, the release of histamine and tumor necrosis factor-alpha was significantly inhibited by resveratrol at a concentration of 100 microM; IgE-mediated release of leukotrienes and prostaglandin D (2) was also strongly suppressed at concentrations of 100 and 10 microM. Also, A23187-mediated release of histamine and leukotrienes release was strongly reduced by resveratrol at concentrations of 100 and 10 microM, respectively. Resveratrol exhibited its behavior without a significant cytotoxic activity against mast cells. In conclusion, resveratrol is a potent non-selective inhibitor of mediator release from activated mast cells and deserves further investigation of its biological modulations.     

Progesterone inhibits mast cell secretion

Mast cells are involved in allergic reactions, where they secrete numerous vasoactive, inflammatory and nociceptive mediators in response to immunoglobulin E (IgE) and antigen. However, they have also been implicated in inflammatory conditions, such as painful bladder syndrome/interstitial cystitis (PBS/IC), irritable bowel syndrome (IBS) and migraines, all of which occur more often in women and are exacerbated during ovulation, but are suppressed during pregnancy. Mast cells express high affinity estrogen receptors and estradiol augments their secretion, while tamoxifen inhibits it. Here we report that progesterone (100 nM), but not the structurally related cholesterol, inhibits histamine secretion from purified rat peritoneal mast cells stimulated immunologically or by substance P (SP), an effect also documented by electron microscopy. These results suggest that mast cell secretion may be regulated by progesterone and may explain the reduced symptoms of certain inflammatory conditions during pregnancy.     

A novel source of mast cells: the human placenta.

The presence of moderate amounts of histamine in the human placenta was confirmed (0.72 +/- 0.10 microgram/g wet weight), and the hitherto unknown storage site of this biogenic amine was elucidated. Mast cells were identified by their characteristic morphology, staining reactions and secretory activity measured in terms of histamine release. Human placental tissue contains 7.6 x 10(5) mast cells/g wet weight, identified by staining with toluidine blue or alcian blue, and these cells were positive for chloro-acetate-esterase. Light microscope studies of placental tissue stained with HRP-conjugated anti-human IgE demonstrated cells with a typical 'halo' effect indicating cell-bound IgE, and electron microscopy revealed cells containing membrane-bound electron dense granules. A single mast cell was calculated to contain approximately 1 pg of histamine. Enzymatic digestion of placental tissue with collagenase (1.5 mg/ml) yielded viable cell suspension. containing mast cells in a purity of 0.6% which exhibited a low spontaneous output of histamine (12%). Placental mast cells released histamine in a concentration dependent manner upon challenge with anti-human IgE and the calcium ionophore A23187. Also, unlike other human mast cells so far studied. with the exception of skin, those dispersed from human placenta were responsive to the polybasic secretagogue compound 48/80. These findings represent a novel source of human mast cells and, since placentas are readily available in quantity, such tissue is proposed as an ideal source of mast cells for biochemical and pharmacological use.     

Confocal laser scanning microscopy to study molecular mechanism of mast cell activation

In the immune system, mast cells are a key cell type in the pathogenesis of immunoglobulin E (IgE)-dependent hypersensitivity reactions. Engagement of the high-affinity IgE receptors by multivalent antigens initiates the downstream activation of signal-transducing enzymes and evokes degranulation and cytokine production via an increase in the intracellular Ca2+ concentration. In addition, mast cells also play a prominent role in non-IgE-mediated hypersensitivity reactions. Mast cells are closely apposed to nerves in vivo and are likely to be regulated functionally by nerves. However, the molecular mechanisms for mast cell activation in an IgE-dependent and -independent manner have not been fully clarified. Confocal laser scanning microscopy has played an essential role in cell biology by allowing visualization of specific intracellular signaling molecules with high spatiotemporal resolution in living cells. We have studied intracellular movements of Ca2+ using a specific fluorescent probe and several types of signaling molecules using derivatives of green fluorescent protein in a living single mast cell using a microscopic strategy. We here describe our imaging analysis of the calcium signals to the nucleus, the movement of secretory granules in the degranulation process, and the nucleocytoplasmic shuttling of mitogen-activated protein kinase in mast cells. Further, we demonstrate that direct communication between mast cells and nerves occurs. These findings provide useful information from a new perspective to understand the molecular mechanisms of allergic reaction and inflammation.     

间质性膀胱炎肥大细胞雌激素受体的表达及意义

目的:检测雌激素受体在间质性膀胱炎(IC)患者膀胱黏膜肥大细胞上的表达情况,探讨雌激素通过调控肥大细胞的激活在IC发病机制中的作用及其临床意义。方法:IC患者膀胱黏膜活检组织标本18例,12例正常膀胱黏膜活检组织标本作为对照组。分别采用0.5%甲苯胺蓝及抗人肥大细胞类胰酶抗体标识组织标本肥大细胞;多克隆抗人雌激素受体抗体标识标本膀胱黏膜上雌激素受体;分析2组标本在肥大细胞数目、脱颗粒肥大细胞比例及雌激素受体表达方面的差异。结果:IC组织标本中肥大细胞密度4.38(3.96,5.29)个/HP,明显高于对照组的2.30(2.26,2.40)个/HP,差异有统计学意义(Z=2.561,P<0.05);IC组织中75.3%的肥大细胞呈不同程度脱颗粒状态,对照组未见明显脱颗粒肥大细胞存在;IC组织标本中雌激素受体阳性肥大细胞(ER+)3.48(3.16,3.69)个/HP,对照组ER+0.82(0.79,0.86)个/HP,2组比较差异有统计学意义(Z=2.988,P<0.05);IC组织中肥大细胞数与ER+数呈正相关(r=0.884,t=3.870,P<0.01)。结论:雌激素可能通过其在肥大细胞上的受体表达来调控肥大细胞脱颗粒,产生IC特有的临床症状。