Hyperplastic and metaplastic lesions in the reproductive tract of male rats induced by neonatal treatment with diethylstilbestrol

Summary Newborn male Wistar rats were castrated on the day of birth (=day 1) and treated daily with diethylstilbestrol (DES) from days 1 to 30; 1 g for the first 10 days of life, 2 g for the next 10 days and 4 g for the last 10 days. The animals were autopsied at 30, 90 and 270 days of age. The epithelium of the coagulating glands (CGs) and ejaculatory ducts (EDs) underwent metaplastic transformation in all DES-treated rats. These pathological changes were more marked with age. The most striking changes were found in the periurethral regions of the CGs and EDs and associated regions of the dorsal urethral wall. The normal transitional epithelial lining almost disappeared and large papillary epithelial outgrowths occurred near the opening of the EDs and CGs. This type of neoplastic change was most marked in the group of rats sacrificed at 9 months of age.     

Isolated polyarteritis nodosa of the male reproductive system associated with a germ cell tumor of the testis: a case report

Polyarteritis nodosa (PAN) presents mostly as a systemic disease with poor prognosis, rarely in an isolated form with a usually favorable outcome. Both forms may affect the male reproductive system and both forms have been associated with malignancies. We describe for the first time the occurrence of isolated PAN in the reproductive system combined with a mixed germ cell tumor of the testis in a 21-year-old man presenting with symptoms of chronic epididymitis. Two years after surgery he is without evidence of recurrence of either the tumor or PAN.     

抑制EGFR信号通路对骨折愈合早期骨内、外膜来源干细胞增殖、分化能力影响的研究*

目的 探索抑制表皮生长因子受体（EGFR）信号通路对骨折愈合早期骨内、外膜来源SCs增殖、分化能力的影响。方法 将建立右股骨骨折模型成功的42只SD大鼠按随机数字表分为实验组与对照组,实验组给予Gefitinib 100 mg/（kg· d）（溶于0.5%甲基纤维素）灌胃,对照组给予等量0.5%甲基纤维素灌胃。术后1周提取右股骨骨内、外膜区域的SCs并进行体外培养。通过流式细胞术鉴定第3代干细胞表面标志物FITC-CD29、FITC-CD34、FITC-CD45、FITC-CD90; BrdU法检测各组干细胞增殖能力; 成骨诱导及成软骨诱导分化后行von Kossa染色及阿尔新蓝染色,检测干细胞的分化能力。结果 各组SCs表面标志物FITC-CD29、FITC-CD90呈高表达,FITC-CD34、FITC-CD45呈低表达; BrdU法检测发现实验组骨内、外膜SCs增殖能力较对照组显著降低（P<0.01）; 成骨诱导分化并染色后,实验组骨内、外膜SCs的矿化结节较对照组更多,成软骨诱导分化并染色后,实验组SCs的淡蓝色颗粒较对照组更多。结论 在骨折愈合早期,抑制EGFR信号通路能抑制骨内、外膜来源SCs的增殖,促进其成骨、成软骨的分化。     

妊娠期高血压疾病胎盘组织中CASR和EGFR的相关研究

目的探讨在妊娠期高血压疾病患者胎盘组织中细胞外钙受体（CASR）和表皮生长因子受体（EGFR）的表达情况及其相互关系。方法通过免疫组织化学方法检测妊娠期高血压疾病患者64例（妊娠期高血压组21例,子痫前期轻度组23例,重度组20例）及健康足月孕妇20例（对照组）胎盘组织中CASR和EGFR蛋白的表达情况。结果 （1）妊娠期高血压疾病患者胎盘组织中CASR在妊娠期高血压组表达水平为59.0532±8.039,子痫前期轻度组患者中为64.3623±3.7278,两组比较,差异有统计学意义（P〈0.0001）;CASR在子痫前期重度组患者中表达为112.2831±6.2060,与妊娠期高血压组比较,差异有统计学意义（P〈0.0001）,与子痫前期轻度组比较,差异有统计学意义（P〈0.0001）。CASR的蛋白在妊娠期高血压组患者胎盘组织中表达水平为59.0532±8.039,对照组为54.8585±4.3035,两组比较,差异无显著性（t=0.08,P=0.7759）。（2）妊娠期高血压疾病患者胎盘组织中表皮生长因子受体（EGFR）在妊娠期高血压组表达水平为56.174±3.1020,子痫前期轻度组患者中为78.6844±2.6713,两组比较,差异有统计学意义（t=18.73,P=0.0001）;EGFR在子痫前期重度组患者中表达为94.2090±6.8352,与子痫前期轻度组比较,差异有统计学意义（t=11.37,P〈0.0001）。（3）胎盘组织中CASR和表皮生长因子受体（EGFR）表达量呈正相关关系（r=0.352,P〈0.05）。结论妊娠期高血压疾病患者胎盘组织中CASR的蛋白表达升高和EGFR激活及过度增高,可能在妊娠期高血压疾病发生发展中起重要作用。     

Prenatal ACTH and corticosterone: effects on reproduction in male mice

Injections of ACTH or corticosterone into late pregnant mice from Days 14 to 21 did not alter testes weights or feminine copulatory behavior of male offspring in adulthood. However masculine copulatory behavior was significantly reduced. Because ACTH does not cross the placenta, alterations in sexual behavior of male offspring were probably mediated by maternal corticosteroid secretion.     

乳腺癌MDA-231细胞中IL-8与表皮生长因子受体信号通路的关系

目的 探讨IL-8对乳腺癌细胞增殖和侵袭的影响以及IL-8信号通路与表皮生长因子受体(EGFR)信号通路之间的关系.方法 采用ELISA方法检测乳腺癌MDA-231细胞IL-8的分泌及rhEGF、anti-EGFR对IL-8分泌的影响;免疫细胞化学方法检测其膜受体CXCR1和CXCR2的表达;MTT和matrigel invasion(基质凝胶侵袭)方法分析rhIL-8及anti-IL-8对癌细胞增殖和侵袭的影响;Western blot分析rhlL-8及anti-IL-8对EGFR活化的影响.结果 乳腺癌细胞可分泌大量IL-8,其细胞膜表面表达CXCR1和CXCR2两种受体.rhlL-8及其中和抗体对乳腺癌细胞的增殖无明显影响,但可影响其侵袭能力:rhIL-8可提高癌细胞的侵袭活性(P＜0.05),其中和抗体则对癌细胞的侵袭有抑制作用(P＜0.05).rhEGF及anti-EGFR均明显抑制了MDA-231细胞IL-8的分泌(P＜0.05,P＜0.01);未发现rhIL-8对EGFR的酪氨酸磷酸化有任何影响,相反anti-IL-8却诱导了EGFR活化.结论 IL-8不是乳腺癌自分泌的促细胞生长因子,IL-8主要通过促进癌细胞的侵袭而影响肿瘤的发展.乳腺癌中G蛋白耦联受体(GPCR)介导的IL-8信号通路与EGFR信号通路之间并无cross-talk,而是竞争抑制的关系.     

Relation of elastosis to biochemical and immunohistochemical steroid receptor findings,Ki-67 and epidermal growth factor receptor (EGFR) immunostaining in invasive ductal breast cancer

We studied 1073 cases of invasive ductal breast cancer, NOS for their elastic content (DEL, ductal + periductal elastosis; TEL, tumour elastosis) and compared the findings with the results of biochemical and immunohistochemical steroid hormone receptor examination. Tumours of patients up to 50 years of age and older were examined separately. In a number of tumours elastosis was also examined in relation to Ki-67 and epidermal growth factor receptor (EGFR) immunostaining. Sensitivity and specificity of DEL and TEL for predicting the receptor, Ki-67 and EGFR findings were estimated. Sensitivity of DEL and TEL for oestrogen and progesterone receptors is dependent on the degree of tumour differentiation and the degree of elastosis, increasing from DEL 1° and TEL 1° to DEL 3° and TEL 3°. It was more evident in grade 1 (G1) and G2 than in G3 carcinomas. Elastosis is a useful predictor of positive receptor findings particularly in G1 and G2 tumours with moderate and high-grade elastosis. It is a similarly useful predictor of negative receptor values in G3 carcinomas. The predictive value of DEL and TEL for the results of Ki-67 and EGFR immunostaining gradually decreases with increasing elastosis, consistent with the assumption that Ki-67 and EGFR identify the degree of tumour proliferation and invasion, while elastosis correlates with the degree of differentiation of breast cancer. Elastosis is a poor predictor of Ki-67 and EGFR findings in any individual breast cancer. Moderate and high-grade elastosis points to positive steroid hormone receptor assays in G1 and G2 carcinomas. In contrast, the lack of elastosis in G3 carcinomas may indicate a negative receptor assay. Both findings have a high degree of reliability.     

表皮生长因子对新生大鼠高氧损伤肺组织EGFR及EGF mRNA表达的影响

目的： 探讨外源性表皮生长因子（EGF）对新生大鼠高氧损伤肺组织EGFR及EGF mRNA表达的影响。方法： 取胎龄21 d剖宫产出生的新生Sprague dawley （SD）大鼠持续吸入95％的O2制作未成熟肺高氧损伤模型，随机分为高氧表皮生长因子（EGF）组和高氧生理盐水（NS）组，另设空气NS对照组；所有组按给药（或NS）时间分为3个亚组，即：a亚组(1-3 d)、b亚组(4-6 d)、c亚组(1-6 d)；各亚组均于生后3、7、14 d分批处死取肺组织。应用免疫组化观察各组肺组织EGF-R的表达，RT-PCR方法检测EGF-mRNA的表达。结果： EGF mRNA的表达随着日龄增加而递增，生后7、14 d高氧组EGF-R及EGF mRNA的表达高于空气对照组，14 d EGFa和c亚组EGF-R的表达均明显高于相应的高氧NS组(P＜0.05)，14 d时EGF组内源性EGF mRNA的表达较NS组明显增加(P＜0.01)。结论： 早期应用EGF可促进肺泡上皮细胞EGF-R的表达，改善高氧所致肺发育受阻，对未成熟肺高氧损伤有一定的保护作用。     

Assessment of epidermal growth factor receptor (EGFR) expression in primary colorectal carcinomas and their related metastases on tissue sections and tissue microarray

Metastatic colorectal carcinomas (CRC) resistant to chemotherapy may benefit from targeting monoclonal therapy cetuximab when they express the epidermal growth factor receptor (EGFR). Because of its clinical implications, we studied EGFR expression by immunohistochemistry on tissue sections of primary CRC (n=32) and their related metastases (n=53). A tissue microarray (TMA) was generated from the same paraffin blocks to determine whether this technique could be used for EGFR screening in CRC. On tissue sections, 84% of the primary CRC and 94% of the metastases were EGFR-positive. When matched, they showed a concordant EGFR-positive status in 78% of the cases. Moreover, staining intensity and extent of EGFR-positive cells in the primary CRC correlated with those observed in the synchronous metastases. On TMA, 65% of the primary CRC, 66% of the metastases, and 43% of the matched primary CRC metastases were EGFR-positive. There was no concordant EGFR status between the primary and the metastatic sites. A strong discrepancy of EGFR status was noted between TMA and tissue sections. In conclusion, EGFR expression measured in tissue sections from primary CRC and their related metastases was found to be similar and frequent, but it was significantly underestimated by the TMA technique.     

Relationship between epidermal growth factor receptor (EGFR) mutation and serum cyclooxygenase-2 Level,and the synergistic effect of celecoxib and gefitinib on EGFR expression in non-small cell lung cancer cells

Epidermal growth factor receptor (EGFR) mutations occur mostly in patients with lung adenocarcinoma; such patients are also more likely to express cyclooxygenase-2 (COX-2), indicating a possible relationship between EGFR mutation and COX-2. The COX-2 and EGFR pathways mutually enhance their procarcinogenic effects in different tumor types. Therefore, simultaneous EGFR and COX-2 inhibition may be a promising therapeutic approach for patients with lung adenocarcinoma. We obtained tissue and serum samples from patients with non-small cell lung cancer (NSCLC) to detect the relationship between EGFR mutation and serum COX-2 level. Subsequently, gefitinib was combined with celecoxib to investigate the efficacy of inhibition in vitro in two NSCLC cell lines: HCC827 (del E746-A750) and A549 (wild-type EGFR). The cells were treated with gefitinib or celecoxib alone or with gefitinib plus celecoxib. Cell proliferation and apoptosis were assessed and correlated with expression of COX-2 and phosphorylated (p)-EGFR. The EGFR mutation rate of the high-COX-2 patients was significantly higher than that in the low-COX-2 patients. Multivariate analysis showed that high COX-2 levels were independently associated with EGFR mutation. Celecoxib and gefitinib inhibited cell growth in both cell lines. At sufficiently high concentrations, celecoxib plus gefitinib significantly mutually enhanced their anti-proliferative and apoptotic effects in both cell lines. At low concentrations, the combination had no additional effects on A549 cells. There was increased down regulation of COX-2 and p-EGFR when both cell lines were treated with high-concentration celecoxib plus gefitinib compared to either agent alone. This study demonstrates that high serum COX-2 levels may indicate EGFR mutations and that the efficacy of combined celecoxib and gefitinib is significantly greater in NSCLC cells with EGFR mutations; at high concentrations, the combination is efficacious in wild-type NSCLC cells.     

EGFR靶向抗体融合蛋白激活免疫细胞杀伤肝癌细胞Huh7的研究

目的 探究靶向表皮生长因子受体(EGFR)的抗体西妥昔单抗(Cetuximab)与MHC-I(major histocompatibility complex-I)相关抗原分子MICB (MHC class I-related chain molecules B)的融合蛋白对肝癌细胞Huh7的杀伤效果.方法 通过重叠PCR(polymerase chain reaction)的方法通过柔性肽G4S将西妥昔单抗重链C末端及人MICB胞外区基因连接,构建成工程载体,并通过毕赤酵母进行表达;流式检测融合蛋白对Huh7细胞的结合能力;噻唑蓝(MTT)法检测Huh7细胞的增殖抑制;通过抗体依赖的细胞介导的细胞毒作用(ADCC)实验验证融合蛋白是否能激活NK细胞杀伤肿瘤细胞.结果 通过基因工程手段和毕赤酵母表达,成功获得融合抗体蛋白,经Western blot鉴定结果说明融合蛋白Cetuximab-MICB表达及装配正确.流式细胞术检测Cetuximab-MICB与肝癌细胞Huh7的结合率为72.8％,与母体西妥昔单抗结合率(75.5％)相当.细胞增殖抑制实验,与PBS组对照相比,西妥昔单抗、融合蛋白组对肝癌细胞Huh7均有抑制作用,且抑制作用呈浓度依赖性.在蛋白浓度为400 nmoL/L时,Cetuximab-MICB融合蛋白组抑制率(66.09％)强于西妥昔单抗组(56.54％).将免疫细胞PBMC(peripheral blood mononuclear cell)/NK92和肿瘤细胞Huh7共孵育,通过检测LDH(lactate dehydrogenase)的释放,发现融合蛋白比西妥昔单抗更有效地介导NK细胞对肿瘤细胞的特异性杀伤,以PBMC为效应细胞,效靶比为100:1时Huh7细胞裂解率可以达到43.58％,优于西妥昔单抗组(37.77％).以NK92细胞为效应细胞,效靶比为10:1时Huh7细胞裂解率可以达到61.29％,明显优于西妥昔单抗组(40.54％).结论 采用毕赤酵母表达体系成功构建并表达了Cetuximab-MICB融合蛋白.Cetuximab-MICB融合蛋白具有良好的体外抗肿瘤活性,有效的抑制肿瘤细胞Huh7的增值,并能进一步通过NKG2D途径激活NK细胞加强免疫系统对肿瘤的免疫监视,为抗肿瘤治疗提供了新的思路,有潜在的临床应用前景.     

Phenylbutenoid dimers isolated from Zingiber purpureum exert neurotrophic effects on cultured neurons and enhance hippocampal neurogenesis in olfactory bulbectomized mice

Trans-3-(3′4′-dimethoxyphenyl)-4-[(E)-3″,4″-dimethoxystyryl]cyclohex-1-ene (Comp.1) and cis-3-(3′4′-dimethoxyphenyl)-4-[(E)-3″,4″-dimethoxystyryl]cyclohex-1-ene (Comp.2), phenylbutenoid dimers, have been isolated as neurotrophic molecules from an Indonesian medicinal plant, Zingiber purpureum. The aim of this study was to explore the neurotrophic effects of Comp.1 and Comp.2 in vitro and in vivo. Comp.1 (10–30 μM) or Comp.2 (30 μM) significantly induced neurite sprouting in PC12 cells. Comp.1 (0.03–3 μM) or Comp.2 (0.3–3 μM) significantly increased the neurite length and number of neurites in primary cultured rat cortical neurons. Comp.1 (30 μM) and Comp.2 (3–30 μM) also provided significant protection against cell death caused by deprivation of serum. The in vivo effects of both Comp.1 and Comp.2 were evaluated on hippocampal neurogenesis in olfactory bulbectomized (OBX) mice, an experimental depression and dementia animal model. Comp.1 (50 mg/kg p.o.), Comp.2 (50 mg/kg p.o.), or fluoxetine (10 mg/kg i.p.), an antidepressant, were administrated once a day on days 15–28 after OBX. Neurogenesis was assessed by analysis of cells expressing NeuN, a neuronal marker, and 5-bromo-2′-deoxyuridine (BrdU) uptake. Immunohistochemical analysis showed that the number of BrdU/NeuN double-labeled cells in the dentate gyrus was significantly decreased 30 days after OBX. Chronic treatment with Comp.1, Comp.2 or fluoxetine significantly increased the number of BrdU/NeuN double-labeled cells. These results indicate that Comp.1 and Comp.2 have neurotrophic effects, and have the potential for disease modification in depression and dementia.