Immunohistochemical distribution of human epidermal growth factor in salivary gland tumours

Summary Immunohistochemical identification of human epidermal growth factor (hEGF) was carried out in a total of 152 cases of salivary gland tumours, consisting 107 pleomorphic adenomas and their variants, 13 adenolymphomas and 32 adenoid cystic carcinomas. A high percentage of pleomorphic adenomas revealed markedly positive hEGF staining of the luminal surface cells of tubuloductal structures and of modified or neoplastic myoepithelial cells. Clear cells of the tumour showed various reactivities from very slight to strong. Eosinophilic epithelial cells of adenolymphoma gave a positive reaction for hEGF in all the cases, whereas most adenoid cystic adenoma lacked hEGF staining; however some cases showed positive staining of the tumour cells. The immunohistochemical detection of hEGF in most salivary gland tumours suggests this factor to be a possible new marker of salivary glands tumours, and to have a biological role in tumour proliferation.     

改构体aFGF对颈动脉窦损伤的保护作用

目的　探讨损伤大白鼠颈动脉窦压力感受器神经末梢后 ,改构体aFGF的保护作用。方法　将Wistar大鼠随机分成对照组、改构体aFGF1组、改构体aFGF2 组、改构体aFGF3 每组各 10只。改构体组分别经静脉注射 0 14 μg/ml、0 4 3μg/ml、1 30 μg/ml的改构体aFGF(1ml/ 10 0g) ,而对照组注射同剂量的生理盐水。 2 0min后 ,用蘸有 80 %乙醇的棉球轻轻擦拭右颈动脉窦区 ,损伤颈动脉窦压力感受器神经末梢 ,并放置 5min。观察记录各组损伤前后夹闭颈总动脉血压即 :收缩压 (SP)、舒张压(DP)、平均动脉压 (MP)和心率 (HR)的变化。结果　 (1)损伤右颈动脉窦前 ,夹闭右颈总动脉 ,实验组和对照组血压均升高 ,夹闭前后相比差异均有统计学意义 (P <0 0 1) ;(2 )损伤右颈动脉窦后 ,夹闭右颈总动脉 ,改构体aFGF1组、对照组血压不变 ,夹闭前后相比差异无统计学意义 (P >0 0 5 ) ;改构体aFGF2 组、改构体aFGF3 组血压升高 ,夹闭前后相比差异有统计学意义 (P <0 0 5 ) ,夹闭前后两组的血压差分别与对照组的差值相比有统计学意义 (P <0 0 5 )。在本实验中 ,HR均无明显变化。结论　改构体aFGF对颈动脉窦损伤有保护作用 ,并显示一定的量效关系     

aFGF重组腺相关病毒转染内皮祖细胞

目的研究含分泌型人酸性成纤维细胞生长因子(sp-haFGF)基因的重组腺相关病毒(rAAV)感染内皮祖细胞(EPCs)的可行性。方法用PCR法将FGF-4的信号序列与原始的aFGF基因连接,形成sp-haFGF基因。将此目的基因克隆到AAV载体质粒pAAV-IRES-hrGFP中,并与包装质粒(pAAV-RC)和辅助质粒(pHelper)共转染HEK293细胞,获得编码sp-haFGF的重组腺相关病毒。用浓缩的病毒感染体外培养的EPC,荧光显微镜下观察细胞表达绿色荧光蛋白情况,并用RT-PCR和W estern b lot方法鉴定sp-haFGF在EPC中的表达。结果获得含有sp-haFGF的重组腺相关病毒,将其感染EPC后,约20%~30%的细胞出现绿色荧光。用RT-PCR法从被感染细胞中扩增出一条长约560 bp的基因条带,通过W estern b lot检测到被感染细胞中haFGF蛋白的表达。结论编码sp-haFGF的重组腺相关病毒能够有效地感染EPC,并在EPC中表达haFGF,为进一步研究转基因的内皮祖细胞移植促血管新生奠定基础。     

成纤维细胞生长因子受体1配体精细结合位点的确定

目的 确定人成纤细胞生长因子受体1（FGFR1）的配体精细结合位点。方法 合成肽文库筛选,定向点突变,源核细胞重组蛋白质表达及受体－配体结合实验。结果 采用^125I标记的酸性成纤维细胞生生长因子（aFGF),自合成肽文库中筛选到一个五肽序列（WGPGM）,其序列及立体结构和FGFR1细胞外段的一个基序（WISPEKM）相似。为了证明FGFR1的WTSPEKM基序是结合FGF的重要结合,采用定向突变技术将WITSPEKM基序中的P（CCA）突变成A（GCA）,并在原核细胞中表达了野生型和突变型FGFR1细胞外段重组蛋白质,受体－配体结合实验显示突变的GFFR1细胞外段结合aFGF的能力明显降低。结论 FGFR1的WISPEKM基序是配体结合的一个重要部位。     

酸性成纤维细胞生长因子对大鼠肾缺血再灌注损伤的保护作用

目的探讨酸性成纤维细胞生长因子(aFGF)对大鼠肾缺血再灌注损伤的影响。方法32只Wistar大鼠随机分为假手术组、模型组、aFGF低剂量组(8!g/kg)和aFGF高剂量组(16!g/kg),每组8只。以肾缺血再灌注损伤为模型,通过光、电镜观察肾组织病理变化。结果aFGF能明显减轻肾组织水肿、肾小管扩张和破坏,电镜下见肾小管上皮细胞超微结构(微绒毛、线粒体、溶酶体等)损伤较轻或基本正常。结论aFGF对大鼠肾缺血再灌注损伤有一定的保护作用。     

血吸虫病肝纤维化中碱性成纤维细胞生长因子的免疫组化研究

目的　研究血吸虫病肝纤维化时碱性成纤维细胞生长因子 (bFGF)在肝脏的表达情况及初步探讨其与纤维化的关系。方法　采用经皮肤感染日本血吸虫尾蚴法建立小鼠血吸虫病肝纤维化模型。利用免疫组化技术研究bFGF在肝纤维化过程中的表达情况 ,并用VG染色显示胶原纤维的沉积情况。结果　小鼠感染后 6周时 ,出现急性血吸虫卵肉芽肿 ,肝内bFGF阳性细胞的分布与对照组无明显差异 ,VG染色显示肉芽肿内无胶原沉积 ;8周时 ,许多bFGF阳性细胞聚集围绕在虫卵肉芽肿周围 ,且细胞形状由原来的多角形变为长梭形。同时 ,VG染色显示肉芽肿周围出现一些胶原纤维 :1 0周时 ,bFGF阳性细胞和胶原纤维均增多。结论　bFGF与血吸虫病肝纤维化有密切关系 ,可能在该病的发生和发展过程中起重要作用     

bFGF在人胚胎干细胞中自我更新的研究进展

人胚胎干细胞具有自我更新和多向分化的独特生物学特性。维持人和小鼠胚胎干细胞增殖的生长因子不同,白血病抑制因子（LIF）不能维持人胚胎干细胞的生长。目前已经确定了数种维持人胚胎干细胞（hESCs）自我更新的生长因子,其中碱性成纤维细胞生长因子（bFGF）信号系统是人胚胎干细胞自我更新中最重要的调节因素之一。将从bFGF及其受体在人胚胎干细胞中的表达和作用的最新进展进行综述。     

人体正常涎腺组织中成纤维细胞生长因子受体3的表达

目的　探讨成纤维细胞生长因子受体 3 (FGFR3 )在人体正常涎腺组织中的表达情况。方法　正常成人涎腺标本2 4例 ,其中腮腺、颌下腺和舌下腺标本各 4例 ,唇腺、腭腺、磨牙后腺及舌腺各 3例 ,分别用超敏S P免疫组化法检测不同腺体组织中FGFR3的分布。结果　在腮腺的浆液性腺泡的分泌颗粒及其他混合性腺的浆液性腺泡的分泌颗粒中均可见少量的FGFR3的表达 ,在大唾液腺及小唾液腺的小叶内导管和小叶间导管中均可见中等强度的FGFR3的表达 ,其中以颌下腺中表达较强 ,而在舌下腺中表达较弱 ,在间质结缔组织中均无FGFR3的表达。结论　人体正常涎腺的导管系统中及浆液性腺泡中有FGFR3的表达     

Immunohistochemical localization of factor X-like antigen in pancreatic islets and their tumours

Summary The authors have investigated by immunohistochemistry the distribution of factor X-like antigen in normal pancreatic islets and in a series of 46 pancreatic endocrine tumours. It was found that both glucagon-producing (A) cells and pancreatic polypeptide-producing (PP) cells are immunoreactive for the antigen. Benign glucagonomas and PP-omas presented the highest concentrations of immunoreactive material whose intracellular distribution was consistent with localization within cell secretory granules. Some benign insulinomas also presented factor X immunostaining in spite of the absence of the antigen in normal insulin-producing B cells. Although malignant tumours usually exhibited very low or no immunostaining, two of three malignant glucagonomas showed scattered, intensely immunoreactive cells. The factor X-like antigen identified in this study was found to differ from chromogranin A and B. The possible implications of the present findings for coagulative disorders associated with glucagonomas or diabetes are discussed.     

Immunohistochemical localization of thrombomodulin in normal human skin and skin tumours

Thrombomodulin (TM) expression has been investigated in sections of normal human skin, in cultured normal human keratinocytes, and in a variety of skin tumours. TM was present in squamous epithelial cells in the spinous layer of normal epidermis and in the outer root sheath of hair follicles, but was absent in the cells of the basal layer. It appeared to be predominantly localized to the cell membrane and the intercellular bridges in these areas. Cultured normal human keratinocytes demonstrated functionally active constitutive TM expression on their cell surface. Immunoperoxidase staining of skin tumours using anti-human TM antibodies demonstrated a typical cell membrane positivity in tumours with squamous or hair follicle differentiation. Basal cell carcinomas showed TM expression only in areas where incomplete squamoid metaplasia occurred. Sweat gland tumours and lesions of the melanogenic system failed to express TM. The localization of TM by immunostaining in various benign and malignant skin tumours typically correlated with their normal skin element of origin. The physiological significance of TM expression in the epidermis is currently undefined.     

Immunohistochemical and ultrastructural localization of tissue polypeptide antigen in human ovarian tumours

Summary Forty specimens of benign and malignant ovarian tumours were studied for localization of tissue polypeptide antigen (TPA) at light and electron microscopic levels by an indirect immunoperoxidase technique. Of the 30 ovarian carcinomas, 23 (77%) were positive and 7 (23%) were negative for TPA, while of the 10 benign ovarian tumours 3 (30%) were positive and 7 (70%) were negative. Positive reaction did not correlate with the tumour grade. Of the 10 patients with metastasis, 8 (80%) had positive tumours. Staining for TPA was observed at the intraluminal cell surfaces and peripheral cell membranes. The ultrastructural localization of TPA revealed electron-dense reaction products at the cell surface and microvillous surfaces. These results provide confirmatory and supplementary evidence to support the previous findings of TPA in the serum and suggest that testing for TPA in ovarian tumors has a limited prognostic importance and a poor diagnostic value. The surface property of TPA suggests that the cell membrane is involved in secretion and probably synthesis of TPA.     

Frequent expression of 75 kDa nerve growth factor receptor and phosphotyrosine in human peripheral nerve tumours: an immunohistochemical study on paraffin-embedded tissues

One hundred and three benign, and 10 malignant peripheral nerve tumours were examined immunohistochemically for expression of 75 kDa nerve growth factor receptor (NGFR). In benign tumours NGFR was demonstrated at 61% in neurinoma, 71% in neurofibroma, 93% in neurofibromatosis and 90% in traumatic neuroma. Malignant neurogenic tumours were 100% positive for NGFR. Phosphotyrosine-immunoreactivity was detected in 76% of NGFR-positive tumours but the frequency of immunostained tumour cells was low. These results suggest that both benign and malignant peripheral nerve tumours express 75 kDa NGFR. The receptor seems to serve as growth signal transduction of the tumour cells in terms of phosphorylation of the tyrosine residue of the receptor or the target protein of the NGFR protein tyrosine kinase.     

An immunohistochemical study of basic fibroblast growth factor in the developing chick

An antiserum against basic fibrobrast growth factor (bFGF) was characterized by immunoblot experiments and used to investigate immunohistochemically the appearance of bFGF-like immunoreactivity in the developing chick. Crude homogenates of chick embryos at every developmental stage, when subjected to immunoblotting with the use of bFGF antiserum, exhibited a main band with the same molecular weight (18 kDa) as bovine bFGF. With immunohistochemistry, bFGF immunoreactivity (bFGF-IR) was detected exclusively in intracellular components of various tissues at different stages of development; bFGF-IR appeared initially on embryonic (incubation) day 3 (E3) in the myotome, on E12 in the spinal cord and ganglia, on E8 in chondrocytes and osteoblasts of the vertebrae, and on E10 in the esophageal epithelium. Immunoreaction products were present either in the cytoplasm or in the nuclei, depending on the types of individual bFGF-containing cells; developing chondrocytes and cells in the stratum basale of the esophagus exhibited intense immunoreactions exclusively within the nuclei, and the other cells mainly within the cytoplasm. Moreover, bFGF-IR was observed in discrete regions of these tissues at different stages; the epithelium of the esophagus containd bFGF-IR in all layers on E10 to E18 with a superficial-to-basal gradient, but it began to exhibit bFGF-IR only in the stratum basale after E20; and bFGF-IR was more abundant in hypertrophic chondrocytes than in proliferating ones. As chicks aged, bFGF-IR decreased or disappeared in the muscles, vertebrae and esophageal epithelium, but neuronal bFGF increased in intensity until the perinatal period and thereafter remained unchanged. These findings suggest that bFGF not only plays a pivotal role in regulating cell proliferation and differentiation in developing chick tissues, but also acts as a non-mitogenic mediator in nervous tissue.     

不同浓度肝细胞生长因子和成纤维细胞生长因子 对大鼠肝干细胞的增殖调控

目的：初步探讨不同浓度的肝细胞生长因子(hepatocyte growth factor,HGF)和成纤维细胞生长因子(fibroblast growth factor, FGF)对大鼠肝干细胞增殖的调控作用。方法：先将大鼠肝胎细胞分别种植A、B培养板上进行肝细胞的体外培养。A组：培养液中含有HGF,浓度分为别1,5,10,20,40,80及100 ng/mL。B组：培养液中含有FGF,浓度分别为1,5,10,20,40,80及100 ng/mL。对照组不加细胞因子。由A、B两组剂量效应获得HGF和FGF的最佳浓度。再将细胞接种于C组培养板进行培养,培养液中含有最佳浓度HGF联合上述各浓度FGF, 并用四甲基偶氮唑盐比色法(mononuclear cell direct cytotoxicity assay, MTT)分别测两组的光密度值（optical delnsity, OD）,检测出HGF与FGF的最佳浓度组合。结果：HGF和FGF浓度与肝干细胞增殖效应呈剂量依赖性（dose-dependent）。当HGF浓度为20 ng/mL,FGF浓度为10 ng/mL时,为最佳浓度组合。结论：HGF和FGF均能对肝干细胞的增殖起促进作用,并且两者有一定协同作用。     

Purification and characterization of fibroblast growth factors from the opossum, Monodelphis domestica, brain.

An extract from the brain of the opossum Monodelphis domestica was fractionated by heparin affinity chromatography. A major peak of mitogenic activity (heparin binding growth factor 2, HBGF-2) eluted from heparin-Sepharose between 1.7 and 2.0 M NaCl. Antisera specific for bovine bFGF detected four polypeptides of 17.5–23 kDa in opossum brain HBGF-2 preparations. Opossum brain heparin binding growth factor 1 (HBGF-1), a minor peak of activity, eluted from heparin-Sepharose at 1.1 NaCl and contained a 16.2 kDa protein that cross-reacted with antiserum against bovine aFGF.     

大鼠肠缺血-再灌注损伤中FGFR2表达的规律

目的观察酸性成纤维细胞生长因子受体2（acid fibroblast growth factor receptor,FGFR2）在肠缺血-再灌注损伤中表达规律。方法以大鼠肠系膜上动脉（SMA）夹闭造成肠缺血-再灌注损伤模型,并将动物随机分为假手术组（C组）、生理盐水对照组（R组）、改构体aFGF治疗组（F组）。除假手术组外,其余各组动物均于缺血45min后于2、6、12、24h活杀,取小肠组织标本,免疫组化和RT—PCR检测FGFR的表达规律。结果在正常大鼠,FGFR分布在小肠绒毛上皮细胞的肠腔侧、侧壁和小肠隐窝朝向隐窝腔的-侧的细胞膜上。缺血和再灌注的初期,FGFR的表达未发生明显变化,随着再灌注时间的延长逐渐增强。FGF使FGFR的表达有明显的增强和提前表达。缺血和再灌注使FGFRmRNA的表达迅速增加,在再灌注后6h达到高峰。F组FGFRmRNA的表达较R组相应时间点显著增加（P〈0．05）。结论FGFR在肠缺血-再灌注损伤的修复中起积极作用。     

碱性成纤维细胞生长因子对体外培养人牙周膜细胞的作用

背景：碱性成纤维细胞生长因子是具有多功能的细胞生长因子,对来源于中胚层及神经外胚层的细胞有明显的促进增殖作用。 目的：观察碱性成纤维细胞生长因子对体外培养人牙周膜细胞的作用。 方法：将第5代人牙周膜细胞,以1×108 L-1的浓度分别接种到96孔板,随机分成4组,分别加入含0,1,10,100 μg/L的碱性成纤维细胞生长因子、体积分数为15%胎牛血清的α-MEM培养基进行培养。在第1,3,5,7天测定细胞的增殖情况,在第1,7天检测碱性磷酸酶活性。 结果与结论：4组之间人牙周膜细胞增殖情况的差异有显著性意义(F=6.586,P=0.024),随着碱性成纤维细胞生长因子质量浓度的增大,吸光度值均增大,其中100 μg/L碱性成纤维细胞生长因子组的吸光度值均大于其他组(P < 0.05);碱性成纤维细胞生长因子各组的碱性磷酸酶活性均低于对照组(P=0.000),浓度越大,活性越低(P < 0.05)。结果显示碱性成纤维细胞生长因子在1-100 μg/L范围内质量浓度越高,促进人牙周膜细胞增殖和抑制碱性磷酸酶活性的作用越强。     

Purification and characterization of fibroblast growth factors from the opossum, Monodelphis domestica, brain

An extract from the brain of the opossum Monodelphis domestica was fractionated by heparin affinity chromatography. A major peak of mitogenic activity (heparin binding growth factor 2, HBGF-2) eluted from heparin-Sepharose between 1.7 and 2.0 M NaCl. Antisera specific for bovine bFGF detected four polypeptides of 17.5–23 kDa in opossum brain HBGF-2 preparations. Opossum brain heparin binding growth factor 1 (HBGF-1), a minor peak of activity, eluted from heparin-Sepharose at 1.1 NaCl and contained a 16.2 kDa protein that cross-reacted with antiserum against bovine aFGF.     

Immunohistochemical distribution of chromogranins A and B and secretogranin II in neuroendocrine tumours of the gastrointestinal tract

The aim of the present study was to investigate immunohistochemically the distribution of chromogranin A, chromogranin B, and secretogranin II in a series of 152 neuroendocrine tumours of the gastrointestinal tract. Tumour tissues from 25 argyrophil gastric carcinoids, 18 gastrin and 5 somatostatin-producing tumours, 4 gangliocytic paragangliomas, 49 classical argentaffin and 2 L cell appendiceal carcinoids, 27 classical ileal carcinoids, 17 rectal carcinoids, and 5 poorly differentiated neuroendocrine tumours of the stomach and rectum were immunostained with antibodies against chromogranin A, chromogranin B, and secretogranin II. Chromogranin A was the major granin expressed in gastric carcinoids and in serotonin-producing carcinoids of the appendix and the ileum. In contrast, strong chromogranin B and secretogranin II immunoreactivity was found in rectal carcinoids, in which chromogranin A was rarely expressed. Since chromogranin A is a widely used marker for neuroendocrine differentiation, it is of diagnostic importance that some gastrin-producing tumours, gangliocytic paragangliomas, poorly differentiated neuroendocrine carcinomas, and appendiceal L cell carcinoids completely lacked chromogranin A positivity. It is concluded that the various neuroendocrine tumours of the gastrointestinal tract show distinctly different patterns of granin expression, probably reflecting their histogenetical origin.