表皮生长因子酶联免疫吸附检测方法的建立

目的建立酶联免疫吸附法(ELISA)检测人血清(浆)或尿液中表皮生长因子的含量。方法以rhEGF为包被抗原，表皮生长因子(epidermal growth factor，EGF)为竞争的抗原，两者与一定量的抗EGF多抗反应。以抑制率为纵坐标，EGF的对数值为横坐标建立标准曲线。结果实验结果表明，理想的包被抗原浓度为1μg／ml，抗EGF多抗工作浓度为1:100000，酶标二抗工作浓度为1:3000，可测量最适范围为0.25-32ng/ml，最小检测量为0．5ng／ml，批内和批间变异系数分别为3．48％和5．46％。得到回归方程y=-0．1281Ln(X)十0．8116(r=-0.9938)和标准曲线。结论建立快速定量测定EGF含量的酶联免疫吸附试验。     

Zonal differences in ethanol-induced impairments in hepatic receptor binding

We have previously shown ethanol-induced defects in receptor-mediated endocytosis of asialoorosomucoid (ASOR), epidermal growth factor (EGF), and insulin in isolated rat hepatocytes. The present study was undertaken to compare the binding of these three ligands in both Zone 1 (periportal [PP] region) and Zone 3 (perivenule [PV] region) of rat liver. Cells from the PV region of ethanol-fed animals bound significantly less EGF (40% decrease) than did cells from the same area in control rats. EGF binding was decreased to a lesser extent (15–25%) in PP cells from ethanol-fed animals compared to controls. When binding of ASOR was examined, ethanol feeding significantly impaired binding in both PP cells (30–35% decrease) and PV cells (50–55% decrease), again showing a greater ethanol-induced impairment in the PV region. Insulin binding in ethanol animals was decreased by 20–25% in both regions compared to controls. In addition, we found that ASOR receptor recycling was impaired to a greater extent in the PV than in the PP region of liver after ethanol feeding, indicating selective impairment of receptor function in the centrilobular region of the liver.     

表皮生长因子信号通路对前列腺癌细胞DU145黏附和迁移能力的影响

目的 探讨表皮生长因子（EGF）信号通路对雄激素非依赖型前列腺癌细胞株DU145黏附和迁移能力的影响。方法 应用细胞黏附能力测定和划痕实验测定EGF对DU145黏附和迁移能力的作用，应用流式细胞术测定EGF对DU145细胞表面整联蛋白α5和β1表达的影响，应用逆转录-聚合酶链反应（RT-PCR）和Western blot测定EGF对DUl45细胞整联蛋白α5和β1亚基总蛋白和mRNA表达的影响。结果 EGF可促进DUl45对纤连蛋白（Fn）的黏附和迁移作用，同时，Fn的受体a5β1的表达发生了变化，EGF上调细胞表面β1亚基的表达，而丝裂源激活蛋白激酶（MAPK）信号通路抑制剂PD98059则具有相反的作用，这种调节作用主要发生在mRNA水平上。结论 EGF可能通过MAPK信号通路，上调β1的表达，从而促进DU145细胞的侵袭能力。     

ZD1839对胰腺癌细胞的生长抑制作用及机理

目的 探讨ZD1839对胰腺癌细胞的生长抑制作用机理.方法 应用MTT方法检测ZD1839对胰腺癌细胞的生长抑制作用、应用不同的生长因子刺激胰腺癌细胞的生长刺激,并检测ZD1839对不同生长因子作用的影响.应用western blot检测不同生长因子对EGF酪氨酸激酶受体的磷酸化作用,以及ZD1839对EGFR受体磷酸化的影响,并检测ZD1839对EGFR信号的下游MAPK磷酸化的影响.结果 ZD1839呈剂量依赖性抑制胰腺癌细胞的生长,ZD1839阻断EGF对胰腺癌细胞的生长刺激作用,但不阻断对IGF-1的作用.ZD1839抑制了基础的与EGF诱导的EGF受体磷酸化水平与MAPK的磷酸化水平.结论 结果表明,EGF对胰腺癌细胞有生长刺激作用,ZD1839对胰腺癌细胞的生长抑制作用是通过对抑制EGF受体磷酸化而特异性起作用的.     

定坤丹联合炔诺酮治疗月经不调的临床研究

目的探讨定坤丹联合炔诺酮治疗月经不调的临床疗效。方法选取2017年3月—2019年3月在开封市人民医院治疗的月经不调患者92例,根据用药的差别分为对照组(46例)和治疗组(46例)。对照组口服炔诺酮片,2.5mg/次,4次/d;治疗组在对照组基础上口服定坤丹,7 g/次,2次/d。两组患者均经过3个月经周期治疗。观察两组患者临床疗效,同时比较治疗前后两组患者临床症候积分、性激素水平、月经失血图(PBAC)、中华生存质量量表(Ch QOL)、高温相评分(HPS)、匹茨堡睡眠质量指数量表(PSQI)评分以及血清细胞因子水平。结果治疗后,对照组临床有效率为82.61%,显著低于治疗组的97.83%,两组比较差异具有统计学意义(P0.05)。治疗后,两组患者症候积分明显下降(P0.05),且治疗组临床症候积分明显低于对照组(P0.05)。治疗后,两组血清孕酮(P)、雌二醇(E2)水平显著升高(P0.05),黄体生成激素(LH)、促卵泡成熟激素(FSH)水平显著下降(P0.05),且治疗组E2、P、LH和FSH水平明显好于对照组(P0.05)。治疗后,两组PBAC和PSQI评分显著下降(P0.05),Ch QOL和HPS评分显著升高(P0.05),且治疗组PBAC、PSQI、Ch QOL和HPS评分明显好于对照组(P0.05)。治疗后,两组患者血清表皮生长因子(EGF)、血管内皮生长因子(VEGF)水平均明显升高(P0.05),且治疗组EGF和VEGF水平明显高于对照组(P0.05)。结论定坤丹联合炔诺酮片治疗月经不调可有效改善患者临床症状,改善机体性激素水平,提高患者睡眠和生活质量。     

多次小剂量STZ处理建立糖尿病大鼠模型及EGF与Gastrin联合应用对胰岛素分泌的影响

目的通过多次小剂量STZ处理建立糖尿病大鼠模型,观察EGF／Gastrin联合应用对模型大鼠胰岛功能的影响。方法采用35mg·Kg^-1腹腔注射STZ一枸橼酸钠缓冲液,每周1次,连续4周,建立大鼠糖尿病模型。采用生化分析和组织染色的方法观察血糖和胰岛细胞的改变。成模大鼠随机分为三组：Insulin治疗组,EGF／Gastrin联合治疗组及DM对照组。EGF／Gastrin组给予EGF（1μg·Kg^-1）／Gastrin（3μg·Kg^-1）,每日1次皮下注射,连续14日;Insulin组给予长效胰岛素2U／d,连续14日;DM对照组及正常对照组给予枸橼酸钠缓冲液。观察血浆中胰岛素和C肽的改变,评估胰岛细胞功能。结果35只实验组大鼠中有33只血糖值≥11．1mmol·L,占总数的94％。Insulin组大鼠和EGF／Gastrin组大鼠血糖值同DM组相比显著降低。DM组中血清胰岛素及c肽含量显著低于正常对照组（P〈0．05）,而Insulin组及EGF／Gastrin组均显著高于DM组（P〈0．05）。组织学结果观察发现,成模大鼠的胰岛数量明显减少,分布不均匀并出现不同程度的萎缩,出现空泡变性。Insulin免疫组织化学染色结果显示,成模大鼠胰岛内Insulin染色阳性的细胞明显减少;且残存胰岛B细胞Insulin染色强度低于正常对照组,Insulin组及EGF／Gastrin组大鼠胰岛中Insulin表达细胞明显增多,有部分细胞呈强阳性表达。结论EGF和Gastrin联合应用能够促进糖尿病大鼠胰岛功能的恢复。     

Epidermal growth factor partially restores colonic ion transport responses in mouse models of chronic colitis

BACKGROUND & AIMS: Epidermal growth factor receptor (EGFR) activation, which plays an important role in regulating intestinal ion transport, can alleviate clinical symptoms such as diarrhea in patients with ulcerative colitis and promote mucosal restitution in animal models of colitis. Here, we investigate whether EGFR can regulate colonic ion transport in the setting of colitis. METHODS: Distal colon from control mice and mice with colitis was retained for immunohistochemistry or mounted in Ussing chambers. Ion transport responses across the tissues to the calcium agonist carbachol and the adenosine 3',5'-cyclic monophosphate agonist forskolin were measured with or without epidermal growth factor (EGF) pretreatment. RESULTS: EGF pretreatment of normal colonic mucosa inhibited ion transport responses to carbachol and forskolin but potentiated the reduced ion transport responses seen in dextran sulfate sodium (DSS)-treated and mdr1a knockout mouse colon. Ion substitution studies and the sodium transport inhibitor amiloride showed that sodium movement primarily accounted for the potentiating effect of EGF in DSS-treated tissues, despite decreased sodium channel expression. EGF potentiation of transport responses in DSS-treated colon was completely blocked by the cytoskeletal disruptor cytochalasin D and the phosphatidylinositol 3-kinase inhibitor wortmannin, whereas the novel and conventional protein kinase C isoform inhibitor G?6850 and the extracellular signal-regulated kinase inhibitor PD98059 partially reduced EGF effects. EGFR epithelial distribution and transforming growth factor alpha expression were also altered in DSS-treated tissues. CONCLUSIONS: Chronic inflammation uncovers a potentiating effect of EGFR activation on epithelial electrogenic sodium absorption that would be expected to ameliorate diarrheal symptoms associated with colitis.     

Effects of epidermal growth factor on neonatal pancreatic growth in the guinea pig

Summary Conclusion EGF and/or transforming growth factor-α (TGF-α) are likely to be important in the rapid pancreatic growth that occurs in the neonatal guinea pig. Background Rapid pancreatic growth is observed during the neonatal period in the guinea pig. The growth factors that are involved are not known but may include members of the EGF family. Methods Mini-osmotic pumps were implanted on the day of birth for continuous infusion of EGF (30 μg/d). Pancreatic DNA, RNA, and protein contents were determined at 4 and 15 d, along with wet weights of the pancreas, duodenum, jejunoileum, colon, and gallbladder. Pancreatic EGF and TGF-α concentrations were measured in adult controls, in control neonates at 1, 4, 8, and 15 d, and also at d 4 and 15 in guinea pigs receiving either EGF or the cholecystokinin receptor antagonist devazepide (25 nmol/kg/h). Results EGF infusion significantly increased the weight of the stomach and duodenum at 4 d and all the gastrointestinal organs, including the pancreas, at 15 d. Exogenous EGF increased pancreatic DNA, RNA, and protein content at 4 and 15 d. Endogenous EGF and TGF-α concentrations in the pancreas were significantly higher at birth than in adults (P<0.001 andP<0.01, respectively) and declined during the first 2 wk postpartum. At 15 d, EGF concentrations remained significantly higher than adult levels (P<0.01), but TGF-α concentrations had declined to adult values. Infusion of EGF decreased concentrations of endogenous EGF in the pancreas at 4 and 15 d (bothP<0.05) and decreased TGF-α concentrations at 4 d (P<0.001). Devazepide infusion caused a significant decrease in endogenous pancreatic EGF concentrations at 15 d (P<0.05).     

血栓调节蛋白基因结构及突变研究进展

血栓调节蛋白是体内抗凝血系统中重要的辅因子。血栓调节蛋白与凝血酶结合活化蛋白C,在凝血和纤溶之间起着重要的联系作用。血栓调节蛋白生物活性的核心位于表皮生长因子4-6区。血栓调节蛋白基因突变与许多疾病具有相关性。A la25Thr突变与50岁以下男性患心肌梗死有关。M et388位改变与静脉血栓相关。A la455Val突变可能促使静脉疾病和脑梗死发生。血栓调节蛋白基因启动子G-33A突变与动脉粥样硬化显著相关。基因突变会对血栓调节蛋白的表达和生物活性产生影响,导致凝血相关的疾病发生。     

补肾调冲方对卵巢早衰大鼠卵泡发育的影响

目的从卵泡发育调控的旁分泌途径研究补肾调冲方的作用机制。方法将60只SD大鼠随机分为正常组,模型组,补肾调冲方高、中、低剂量组及西药组,每组10只,采用雷公藤多苷灌胃(50mg/kg,14天)建立卵巢早衰模型,分别使用高(1ml/100g)、中(0.5ml/100g)、低(0.25ml/100g)剂量补肾调冲方及倍美力(0.075ml/100g)进行干预,时间为35天,观察各组大鼠各级卵泡及黄体计数和卵巢超微结构改变,检测卵巢卵泡刺激素受体(FSHR)、表皮生长因子(EGF)及缝隙连接蛋白43(Cx43)的mRNA表达。结果模型组各级卵泡数、黄体数明显减少,卵巢超微结构受损,EGF、Cx43mRNA表达显著降低(P<0.05或P<0.01),补肾调冲方中、低剂量组及西药组大、中卵泡数及黄体数明显增加,卵巢超微结构明显改善,EGF及Cx43的mRNA表达显著增高(P<0.05或P<0.01)。结论补肾调冲方可改善雷公藤多苷所致的卵泡发育障碍,其作用机制可能是通过促进卵巢EGF、Cx43mRNA表达这一旁分泌调控途径实现。