肥大细胞的研究进展

肥大细胞(MC)是系统发生学的一种成熟细胞。在哺乳动物,MC分布于除骨、软骨和眼球角膜的结缔组织,并以相当一致方式围绕血管、淋巴管、和神经末梢分布。MC是一种成活时间长的多功能分泌细胞,胞浆内有大量由蛋白多糖基质、肝素组成的高电子密度颗粒。它是哺乳动物组织内组胺的主要储藏地。本文就MC的表型、测定方法、介质等方面的研究现状作一简要介绍。     

肥大细胞与肺动脉高压

肥大细胞是人类速发型过敏反应的起始细胞，它广泛分布于结缔组织、粘膜组织及血管周围。肥大细胞通过释放脂性介质、分泌颗粒介质和细胞因子等来发挥其作用。肥大细胞分泌颗粒内含有多种活性介质，主要包括组胺、肝素、类糜蛋白酶、类胰蛋白酶以及白三烯和多种细胞因子。人肥大细胞主要有结缔组织型肥大细胞（CTMC）和粘膜组织型肥大细胞（MMC）两类。     

肥大细胞脱颗粒的标志

肥大细胞 (mastcell ,MC)是人速发型过敏反应的主要效应细胞。MC激活后 ,预先形成的介质及新合成的介质一起释放。组胺含量可通过荧光分光光度法 ,放射免疫分析法及酶免疫分析法测定。这些分析法在体内已用于血浆及尿内的组胺含量测定 ,在体外可用于测定嗜碱粒细胞及肥大细胞的组胺释放。类胰蛋白酶是MC更具有选择性的标志 ,因为嗜碱粒细胞几乎不含此成分。β 类胰蛋白酶贮存于分泌颗粒中 ,MC激活时 ,被释放至细胞外。α 类胰蛋白酶原以酶原的形式存在 ,它的释放不需要MC的激活 ,所以它的血清水平可以反映体内MC数目。α 类胰蛋白酶原在系统性肥大细胞增生症病人的血清中含量增高 ,而 β 类胰蛋白酶在系统性过敏反应病人血清中含量升高。这些变化可作为MC参与疾病发展的准确的临床指征     

肥大细胞脱颗粒的标志

肥大细胞（mast cell,MC)是人速发型过敏反应的主要效应细胞,MC激活后,预先形成的介质及新合成的介素一起释放。组胺含量可通过蒌光分光光度法,放射免疫分析法及酶免疫分析法测定,这些分析法在体内己用于血浆及尿内的组胺含量测定,在体外可用于测定嗜碱粒细胞及肥大细胞的组胺释放,类胰蛋白酶是MC更具有选择性的标志,因为嗜碱粒细胞几乎不含此成分,β-类胰蛋白酶贮存于分泌颗粒中,MC激活时,被放放至细胞外,α-类胰蛋白酶原以酶原的形成存在,它的释放不需要MC的激活,所以它的血清水平可以反映体人MC数目。α-类胰蛋白酶原在系统性肥大细胞增生症病人的血清中含量增高,而β-类胰蛋白酶在系统性过敏反应病人血清中含量升高,这些变化可作为MC参与疾病发展的准确的临床指征。     

肥大细胞的研究进展

肥大细胞(MC)是系统发生学的一种成熟细胞。在哺乳动物，MC分布于除骨、软骨和眼球角膜的结缔组织，并以相当一致方式围绕血管、淋巴管、和神经末梢分布。MC是一种成活时间长的多功能分泌细胞，胞浆内有大量由蛋白多糖基质、肝素组成的高电子密度颗粒。它是哺乳动物组织内组胺的主要储藏地。本文就MC的表型、测定方法、介质等方面的研究现状作一简要介绍。     

肥大细胞增生症临床病理学观察

目的 探讨肥大细胞增生症的临床病理学特点及诊断.方法 对2例肥大细胞增生症进行临床资料分析、病理形态学观察、免疫表型和特殊染色检测.结果 例1女婴,出生3个小时,出生时全身弥漫性皮疹,临床取腹部皮肤皮疹活检.例2男性,47岁,间断性腹泻7年,体检发现肝脾肿大,临床先后行结肠镜活检、骨髓穿刺活检和肝穿刺活检.组织形态学表现:例1皮肤真皮内有片状单核样细胞浸润.例2结肠黏膜内、骨髓内、肝窦及汇管区内见片状形态较一致的单核样细胞浸润,胞质较丰富,呈细颗粒状.免疫组化显示CD117和CD68(+),CD3、CD20、CD25、CD30、S-100和MPO(-).甲苯胺蓝染色细胞胞质内见紫蓝色颗粒.结论 肥大细胞增生症易误诊为其他白血病/淋巴瘤,以消化道症状为首发时常被误诊为炎症性肠病.利用特殊染色和免疫表型来证实肥大细胞对确定诊断十分重要.     

胃癌癌周肥大细胞密度与淋巴结癌转移的关系

目的：探讨胃癌癌周肥大细胞（ＭＣ）密度与淋巴结癌转移的关系。方法：应用俾士麦棕法检测伴有淋巴结癌转移的２５３例胃癌病例。结果：１￣６ＩＬＮ病例ＭＣ密度高于≥７￣１５ＩＬＮ病便,７￣１５ＩＬＮ病例ＭＣ密度高于〉１５ＩＬＮ病例,Ｐ均〈０．０１。≥５年生存年限的病例ＭＣ密度高于〈５年生存年限病便的ＭＣ密度,Ｐ〈０．０１。５年生存率,１－６ＩＬＮ病例高于７￣１５ＩＬＮ病例,７￣１５ＩＬＮ病例高于〉１５ＩＬ     

肥大细胞促早期胃癌形成及其生物学意义

目的通过分析胃癌、胃癌前病变及胃良性病变组织中肥大细胞数目分布,了解肥大细胞与肿瘤形成及生存期的关系。方法回顾性分析59例胃癌、40例胃癌前病变、39例胃良性病变患者的临床病理及随访资料。采用免疫组化标记肥大细胞特异性类胰蛋白酶,分析肥大细胞在胃癌(高、中、低分化)组织中的数目及分布,并与胃癌前病变、胃良性病变组织中的表达进行比较,同时分析其与临床病理资料及生存期的关系。结果胃癌、胃癌前病变组织中肥大细胞数明显高于胃良性病变(P均<0.05),胃癌前病变组织中肥大细胞数明显高于胃癌(P<0.05)。将胃癌组和胃癌前病变组分别以肥大细胞数中位数(28、43)分组比较,胃癌、胃癌前病变中肥大细胞高水平组和低水平组生存期比较差异无统计学意义(P均>0.05)。肥大细胞数与幽门螺杆菌感染及Ki-67表达强弱有明显相关性(P均<0.05)。结论胃癌、胃癌前病变、胃良性病变组织中肥大细胞差异提示肥大细胞可能参与了早期肿瘤的形成,特别是幽门螺杆菌感染诱发的胃癌;肥大细胞与Ki-67的关系提示其促进肿瘤增殖与进展,但是与肿瘤分化程度无关;同时,肿瘤组织中肥大细胞与患者生存期无明显相关性。     

肥大细胞类胰蛋白酶与疾病

肥大细胞类胰蛋白酶作为肥大细胞特异性标志物可用于肥大细胞的鉴定和分型，类胰蛋白酶主要有α-类胰蛋白酶和β-类胰蛋白酶两种。在系统性肥大细胞增生症病人血清中，α-类胰蛋白酶水平增高；在过敏性疾病中，β-类胰蛋白酶和组胺同时测定能够用于监测过敏反应；类胰蛋白酶还与很多疾病如血栓症、慢性阻塞性肺疾病及肿瘤的发生过程有关联，其抑制剂的开发为炎性疾病的治疗提供了广阔的前景。     

辛伐它汀干预后肥大细胞在肺动脉高压中的变化

目的:建立SD大鼠高肺血流,注射野百合碱所致肺动脉高压模型,并用辛伐它汀干预,观察肺组织中肥大细胞及肥大细胞两种亚型的变化,探讨辛伐它汀干预后肥大细胞在肺动脉高压中的改变及其机制。方法:采用腹主动脉-下腔静脉造瘘,术后第二天治疗组给予辛伐它汀2mg/kg干预,并于术后7天给予治疗组和模型组一次性腹腔注射野百合碱60mg/kg,制造肺动脉高压(pulmonary hypertension)动物模型,正常SD大鼠作为对照组,35天后测定肺动脉压力。取右下肺组织,用抗类胰蛋白酶单克隆抗体和抗类糜蛋白酶单克隆抗体免疫组织化学染色观察实验组与正常组之间肥大细胞数量的变化,HE染色观察肺组织一般病理变化。结果:HE染色显示模型组肺组织中动脉管壁明显曾粗,有新生血管形成。平均肺动脉压力(mPAP)模型组动物较对照组明显升高(P<0.05),模型组较治疗组也明显升高(P<0.05);右心室/(左心室 室间隔)(RV/LV S)模型组较对照组明显增加(P<0.05),模型组较治疗组也明显增加(P<0.05),肥大细胞总数(类胰蛋白酶阳性的肥大细胞)及类糜蛋白酶阳性的肥大细胞模型组较对照组明显增多(P<0.05),肥大细胞在治疗组较模型组明显减少(P<0.05)。结论:腹主动脉-下腔静脉造瘘合并野百合碱腹腔注射成功地建立了肺动脉高压的动物模型,已有文献报道肥大细胞可能参与了肺动脉高压的形成,免疫组织化学染色显示在辛伐它汀干预后肥大细胞的总数治疗组较模型组明显减少,对临床治疗肺动脉高压提供了新的思路。     

自然杀伤细胞在胃癌组织中的表达及临床意义

目的分析自然杀伤（NK）细胞在胃癌组织中的表达及其与胃癌临床病理特征和预后的关系。方法采用免疫组织化学染色法检测107例胃癌患者手术标本中NK细胞的表达,采用SAS9．12统计软件包进行资料分析。结果在胃癌、胃炎及胃息肉组织中均可见NK细胞的表达,但胃癌组NK细胞的表达[（31．74±15．93）个／HPF]明显高于慢性胃炎组[（5．02±2．55）个／HPF]和胃息肉组[（4．84±1．55）个／HPF],差异有统计学意义（P〈0．01）。NK细胞的表达与胃癌患者的性别、年龄、肿瘤部位、肿瘤大小、淋巴结转移、复发、病理分级、组织学类型和浸润深度均无统计学意义（P〉0．05）,但TNM分期越高,NK细胞表达水平有明显下降的趋势,差异有统计学意义（P〈0．05）。与NK细胞低表达组相比,NK细胞高表达组胃癌患者中位生存时间延长14个月,两组生存曲线差异无统计学意义（χ2=2．888,P〉0．05）。多因素COX模型分析显示,与NK细胞低表达组相比,高表达组有降低胃癌死亡风险的趋势（RR=0．68,95％CI=0．41～1．23）。结论NK细胞的高表达与胃癌患者的生存时间呈正相关,可作为预测胃癌生存期的指标。     

Clinicopathological significance of PI3K,Akt and survivin expression in gastric cancer

Background

Gastric cancer is the second leading cause of cancer-related deaths worldwide, and is characterized by invasion and metastasis. Increasing attention is being focused on discovering molecular markers for diagnosis and prognosis. Our objective was to evaluate PI3K, Akt and survivin protein expression in gastric cancer, and their correlations with clinicohistological features and prognosis in patients with gastric cancer.

Methods

Tissue samples were obtained from 70 patients with gastric cancer patients and 20 patients with normal gastric mucosa. The protein levels of PI3K, Akt and survivin were evaluated by immunohistochemistry. Statistical analyses were performed to establish the correlations between their expressions and patients’ clinicopathologic characteristics.

Results

The positive expression rates of PI3K, Akt and survivin were significantly higher in the gastric cancer tissues compared to normal gastric mucosa (P < 0.05). Expression levels of PI3K, Akt and survivin proteins were significantly correlated with TNM stage, differentiation grade, lymph node metastasis and metastases to other organs (P < 0.05). Cooperative relationships were identified between PI3K and Akt, and PI3K and survivin (P < 0.01), suggesting the involvement of the PI3K/Akt/survivin signaling pathway in the tumorigenesis of gastric cancer.

Conclusions

Protein expression of PI3K, Akt and survivin were significantly associated with the development, progression and metastasis of gastric cancer and may have value as diagnostic and prognostic markers in gastric cancer.     

热休克胃癌细胞负载DC-CIK细胞杀伤癌细胞活性的研究

目的研究负载相关抗原的树突状细胞(DC)联合细胞因子诱导的杀伤(CIK)细胞对胃癌细胞的杀伤作用。方法取健康人外周血并分离出外周血单个核细胞(PBMC),经不同细胞因子分别诱导出DC和CIK细胞,热休克方法处理胃癌MKN-28细胞株并制备肿瘤抗原用于负载DC,与CIK细胞共培养,以流式细胞术检测DC、CIK细胞表型,MTS法检测CIK细胞对MKN-28细胞的杀伤作用。结果 CIK与DC共培养后,与单纯CIK细胞相比较,增殖活性明显提高,CD3+CD8+、CD3+CD56+双阳性细胞数增多并且具有更强的杀瘤活性,效靶比为20.0∶1时,负载MKN-28抗原的DC-CIK(Ag-DC-CIK)组对MKN-28的杀伤活性为(57.96±2.23)%,远大于未负载MKN-28抗原的DC-CIK组[(38.02±2.06)%]和单纯CIK组[(29.78±1.84)%],差异具有统计学意义(P<0.05)。结论以热休克凋亡肿瘤细胞抗原负载的DC能促进CIK细胞增殖分化,并提高其对胃癌细胞的杀伤活性。     

The mechanism between epithelial mesenchymal transition in breast cancer and hypoxia microenvironment

Hypoxia microenvironment widely exists in solid tumor tissues, which is mainly due to the rapid growth of cells within the tumor more than the speed of capillary in neoplasm, resulting in tumor tissue hypoxia. In hypoxia, hypoxia inducible factor 1 (HIF-1) is activated and regulate the expression of a series of hypoxia inducible genes, resulting in a series of hypoxia adaptation reaction. Researchs proved that, HIF-1 is closely related to the invasion, metastasis, prognosis of the tumor, and the expression of HIF-1 is higher in metastatic tissues compared with primary cancer tissues. In the evolution process of breast cancer, epithelial mesenchymal transition (EMT) define the characteristics of migration and invasion of breast cancer cells, which can also allow cancer cells to acquire the ability of self-renewing and stemness, so as to promote the generation of breast cancer stem cells. The incidence of EMT cancer stem cells are higher within the resistant to conventional treatment. This review focuses on breast cancer (stem cells), targeting the mechanism between hypoxia and EMT in tumor (stem cells), with the purpose of finding the new therapy to breast cancer.     

人胃癌微环境来源间充质干细胞促进胃癌细胞BGC-823的体外增殖、迁移和促血管生成能力

目的探讨人胃癌微环境来源间充质干细胞(GC-MSCs)能否促进胃癌细胞增殖、迁移及其体外促血管生成能力。方法采用MTT增殖试验和细胞克隆形成试验观察GC-MSCs细胞对胃癌细胞系BGC-823生长的影响;Transwell迁移试验检测GC-MSCs细胞对BGC-823细胞迁移能力的影响;利用管形成试验观察并分析GC-MSCs细胞对BGC-823细胞促血管生成能力的影响;RT-PCR检测GC-MSCs细胞作用BGC-823细胞后其促血管生成相关基因(VEGF、MIP-2和TGF-β1基因)的表达情况。结果 MTT增殖试验和克隆形成试验结果表明,GC-MSCs来源的细胞培养上清可以明显促进胃癌细胞BGC-823的体外生长(P0.05);Transwell迁移试验结果表明,GC-MSCs来源细胞培养上清对BGC-823的迁移能力具有显著促进作用(P0.01);RT-PCR检测结果显示,GC-MSCs来源的细胞培养上清可明显上调促血管生成相关因子(VEGF、MIP-2和TGF-β1基因)在BGC-823细胞中的表达水平;管形成试验结果显示,BGC-823和GC-MSCs细胞共培养上清与BGC-823、GC-MSCs单独培养组相比具有更强的促血管生成能力。结论 GC-MSCs可通过旁分泌的方式促进胃癌细胞增殖、迁移及其促血管生成能力,进而诱导胃癌的恶性转化。     

Cysteinyl cathepsins and mast cell proteases in the pathogenesis and therapeutics of cardiovascular diseases

The initiation and progression of cardiovascular diseases involve extensive arterial wall matrix protein degradation. Proteases are essential to these pathological events. Recent discoveries suggest that proteases do more than catabolize matrix proteins. During the pathogenesis of atherosclerosis, abdominal aortic aneuryms, and associated complications, cysteinyl cathepsins and mast cell tryptases and chymases participate importantly in vascular cell apoptosis, foam cell formation, matrix protein gene expression, and pro-enzyme, latent cytokine, chemokine, and growth factor activation. Experimental animal disease models have been invaluable in examining each of these protease functions. Deficiency and pharmacological inhibition of cathepsins or mast cell proteases have allowed their in vivo evaluation in the setting of pathological conditions. Recent discoveries of highly selective and potent inhibitors of cathepsins, chymase, and tryptase, and their applications in vascular diseases in animal models and non-vascular diseases in human trials, have led to the hypothesis that selective inhibition of cathepsins, chymases, and tryptase will benefit patients suffering from cardiovascular diseases. This review highlights recent discoveries from in vitro cell-based studies to experimental animal cardiovascular disease models, from protease knockout mice to treatments with recently developed selective and potent protease inhibitors, and from patients with cathepsin-associated non-vascular diseases to those affected by cardiovascular complications.     

胃癌患者组织中树突状细胞亚型的表达及临床意义

目的探讨树突状细胞(DC)亚型在胃癌患者组织中的表达水平及其与临床病理特征及预后的关系。方法 202例随访胃癌患者中有140例符合纳入标准,用流式细胞术检测胃癌患者组织中DC亚型;免疫组织化学染色法检测140例胃癌患者癌组织和10例慢性胃炎组织中CD11c+DC的表达水平,并结合临床病理特征进行统计学分析。结果在胃癌组织中单核细胞来源DC(Mo DC)的表达水平为(6.3±6.3)%,而髓系来源的DC(m DC)及浆细胞来源DC(p DC)均不表达;胃癌组织中CD11c+DC表达水平与肿瘤大小、淋巴结转移、病理分级、TNM分期比较差异均有统计学意义(P均0.05);且其表达水平和TNM分期是胃癌患者预后的重要影响因素(P均0.05)。结论胃癌组织中浸润的DC细胞仅为Mo DC;CD11c+DC与胃癌发生和转移有关,可能参与调控肿瘤免疫应答。     

基于数据挖掘分析GPX4在胃癌中的表达及临床意义

目的探索谷胱甘肽过氧化物酶4（GPX4）在胃癌中的表达及临床意义。 方法检索收集Oncomine和基因表达谱动态分析（GEPIA）数据库中GPX4 mRNA在常见肿瘤中的表达情况,并分析其在胃癌中的表达及与患者预后的关系。利用cBioPortal在线分析癌症基因图谱（TCGA）数据库中1178例胃癌样本中的RNA测序数据,分析GPX4基因的突变情况及其与预后的关系。利用String数据库预测与GPX4相互作用的蛋白。 结果Oncomine数据库中共收集了459项关于GPX4基因在常见肿瘤及正常组织中的对照研究,其中22项表达差异具有统计学意义（P<0.0001）。GEPIA数据库分析结果显示GPX4 mRNA在胃癌组织中高表达（P<0.01）,且与不良的总体生存期及无进展生存期明显相关（P<0.05）,但与胃癌的TNM分期无明显相关性（P>0.05）。对TCGA数据库的RNA测序结果分析发现,在1178例胃癌样本中有25例发生GPX4基因变异,总变异率为2.1%。Kaplan-Meier生存曲线分析显示,GPX4基因变异与否,与总体生存期及无进展生存期无明显相关性（P>0.05）。GSR、GSS、GGT1/5/6/7,GSTO2,GRSF1,SOD1/2等蛋白与GPX4有明显的相互作用（P=1.94×10^-7）。 结论利用多种肿瘤基因数据库分析表明,GPX4基因在胃癌组织中变异率低,其mRNA水平高表达与患者不良预后相关,为进一步探究GPX4在胃癌的发生发展中的作用提供了重要理论依据。     

Contrast media are incomplete secretagogues acting on human basophils and mast cells isolated from heart and lung,but not skin tissue

To investigate the mechanisms of anaphylactoid reactions to radiocontrast media, in vitro mediator release induced by three iodinated contrast agents was examined using peripheral blood basophils and mast cells purified from human lung parenchyma, heart, and skin tissues. Three iodinated contrast agents, sodium and meglumine salts of ioxaglic acid, sodium and meglumine salts of ioxithalamic acid, and ioversol, were incubated with basophils purified from peripheral blood and human mast cells isolated and purified from different anatomical sites. Release of preformed (histamine and tryptase) and de novo synthesized mediators (prostaglandin D₂ and leukotriene C₄) into the supernatans was determined at various contrast medium concentrations after incubation for 60 min. Ioxaglate (0.2–0.3 M), ioxithalamate (0.3–0.5 M), and to a lesser extent ioversol (0.3–0.5 M) induced histamine release from basophils in a concentration-dependent manner. All three induced the release of preformed mediators (histamine and tryptase) from human lung, but not from skin mast cells. They also induced histamine and tryptase release from human heart mast cells. However, they did not induce the de novo synthesis of leukotriene C₄ or prostaglandin D₂ from human basophils or any type of mast cell examined. Cross-linking of IgE by anti-IgE induced the release of leukotriene C₄ or prostaglandin D₂ from human basophils or mast cells. Mannitol, an osmotic stimulus, induced the release of histamine from human basophils, but to a lesser extent from mast cells. These results show that different contrast media can differ in their ability to release mediators from enriched preparations of human basophils and mast cells. The three contrast agents examined act on basophils and mast cells as incomplete secretagogues, causing the release of preformed mediators, but not the de novo synthesis of chemical mediators. It may be useful to measure plasma tryptase levels to detect adverse reactions caused by iodinated radiographic contrast materials.