Cellular localization of immunoreactive epidermal growth factor during Wolffian duct differentiation of the fetal mouse

Previous studies from this laboratory indicated a role for epidermal growth factor (EGF) in androgen-dependent male reproductive tract differentiation of the fetal mouse. Expression of an EGF-like protein during Wolffian duct differentiation was indicated from the determinations by radioimmunoassay (RIA) and radioreceptor assay. To further characterize the protein and to assess its role in male sexual differentiation, expression of the protein has been analyzed by Western blot assay and its tissue-specific cellular expression has been determined by immunocytochemical assay in the present study. Western blot analysis of the 18-day fetal male reproductive tract detected an immunoreactive band of the predicted 6-kDa size. Immunocytochemical analysis also detected EGF-specific immunostaining in the Wolffian duct derivatives. At day 18 of gestation, the staining was localized predominantly in the epithelial nuclei of the Wolffian duct derivatives whereas at days 14 and 16 of gestation, the staining was equally distributed in the mesenchymal and epithelial sites of the Wolffian duct derivative. The intensity of the staining increased with progression of differentiation during the 14th–18th days of gestation. Prenatal exposure to the antiandrogen flutamide significantly reduced the immunostaining of the duct. Thus, a role for EGF in Wolffian duct differentiation is indicated.     

同型半胱氨酸对胚胎海马神经元细胞的影响

目的 观察同型半胱氨酸(homocysteine，HCY)对海马神经元细胞分化和增殖的作用机理。方法 采用大鼠胚胎海马神经元细胞进行体外培养。观察不同浓度的HCY(0．01、0．1、1．0、10．0、100．0)mmol／L对细胞分化和增殖的影响。结果 随着剂量的增加．同型半胱氨酸对胚胎海马神经元细胞分化和增殖具有抑制作用，并用流式细胞术分析证明对脂质过氧化(LPO)的影响而使得神经元蛋白质含量增高。结论 同型半胱氨酸可抑制胚胎海马神经元细胞分化和增殖的影响，HCY可能在导致神经管畸形的过程起着重要作用。     

Mucinous adenocarcinoma with neuroendocrine differentiation of the mandibular ramus: report of a case

A rare case of mucinous adenocarcinoma with neuroendocrine differentiation of the mandibular ramus is presented. The patient, an 80-year-old man, was referred to our hospital with chief complaint of swelling and pain in the left buccal mucosa. CT and MRI examination showed an osteolytic tumor mass occupying the upper region of the left mandibular ramus. Macroscopically, the excised tumor was a relatively well-defined, solid mass with diffuse bone resorption, measuring 3 cm x 3.2 cm x 3 cm. Microscopical examination showed that the tumor forming glandular structures with abundant mucous production and high cellular atypia. Immunohistochemical studies demonstrated the positive reactivities for pan-keratin, cytokeratin 7, vimentin,alpha-amylase, alpha-smooth muscle actin, neuron-specific enolase, glial fibrillary acid protein, calcitonin, and somatostatin in tumor cells. These findings suggested that the tumor was originated from heterotopic or misplaced salivary gland in the mandible.     

Induction of Donor-Specific Allograft Tolerance by Short-Term Treatment with LF15-0195 After Transplantation. Evidence for a Direct Effect on T-Cell Differentiation

A 20-day treatment with LF15-0195, a deoxyspergualine analog, induced long-term heart allograft survival in the rat without signs of chronic rejection. LF15-0195-treated recipients did not develop an anti-donor alloantibody response. Analysis of graft-infiltrating cells, IL10, TNFalpha, IFNgamma mRNA and iNOS protein expression in allografts, 5 days after transplantation, showed that they were markedly decreased in allografts from LF15-0195-treated recipients compared with allografts from untreated recipients. Surprisingly, spleen T cells from LF15-0195 recipients, 5days after grafting, were able to proliferate strongly in vitro, when stimulated with donor cells, but had reduced mRNA expression for IFNy compared with spleen T cells from untreated graft recipients. Furthermore, when T cells from naive animals were stimulated in vitro, using anti-CD3 and anti-CD28, LF15-0195 also increased T-cell proliferation in a dose-dependent fashion: however, these cells expressed less of the Th1 -related cytokines, IFNgamma and IL2, compared with untreated cells, suggesting that LF15-0195 could act on T-cell differentiation. In conclusion, we show here that a short-term treatment with LF15-0195 induced long-term allograft tolerance, decreasing the in situ anti-donor response, and we illustrate evidence for the development of regulatory mechanisms.     

胃肠富集kruppel因子GKLF在宫颈鳞癌中的表达及意义

目的　探讨胃肠富集kruppel因子 (GKLF)在宫颈鳞癌组织及正常宫颈组织中的表达及意义。方法　应用半定量的逆转录聚合酶链式反应 (RT -PCR)方法检测 32例宫颈鳞癌组织 (研究组 )中GKLFmRNA的表达强度 ,以 10例正常宫颈组织作为对照 (对照组 )。结果　GKLFmRNA在正常宫颈组织中的表达强度为 0 .76± 0 .15 ,而在宫颈鳞癌中的相对表达强度为 0 .4 2± 0 .19,与正常宫颈组织相比较 ,宫颈鳞癌组织中GKLFmRNA的表达丰度较低 (P <0 .0 5 )。而且临床分期愈晚 ,GKLFmRNA的表达强度愈低 ,差异有显著性 (P <0 .0 5 )。随着病理分级增高 ,GKLFmRNA的表达强度逐渐降低 ,差异有显著性 (P <0 .0 5 )。结论　宫颈鳞癌组织中GKLF表达下调 ,而且GKLFmRNA的表达与宫颈癌的临床分期和病理分级呈负相关 ,提示GKLF可能参与了宫颈癌的发生或进展过程     

小鼠牙髓干细胞培养及诱导分化

目的 :分离、培养小鼠牙髓干细胞并诱导分化 .方法 :采用酶消化培养法获得小鼠的单个牙髓细胞悬液 ,细胞密度调整为 1× 10 4/孔细胞 ,用 2 0 0mL·L -1胎牛血清的α MEM培养液培养 14d ,挑出细胞克隆消化传代 ,TGF β、BMP2 、bFGF细胞因子诱导 ,PCR检测DSPP的mRNA的表达 .结果 :小鼠牙髓细胞呈集落状生长 ,克隆形成率为 1.6 -2 .5个 / 10 4细胞 ,所形成的集落细胞结合紧密 ,细胞体积小、胞核大 ,经细胞因子刺激后的细胞表达DSPP的mRNA .结论 :培养的小鼠牙髓集落状生长的细胞具有干细胞增殖快等特性 ,细胞因子可诱导小鼠牙髓干细胞定向分化     

骨髓间充质干细胞体外诱导为神经细胞的研究

本实验观察了大鼠MSCs向神经细胞方向的诱导分化情况,以期为MSCs在神经移植领域的临床应用提供理论基础。用含10 ng／ml bFGF+20％FBS的DMEM对MSCs进行预诱导24 h后,以含200μmol／L的BHA+2％DMSO的无血清DMEM对MSCs进行诱导,观察诱导后细胞的光、电镜形态学变化,通过免疫组织化学法对诱导后细胞进行神经细胞表型及神经递质合成酶鉴定。结果显示:MSCs经BHA和DMSO诱导后,80％以上的细胞表现出神经元样形态,胞浆内可见较多Nissl体,并表达nestin、NSE、NF、MAP、SYN,部分诱导后的细胞表达ChAT、TH、GAD;电镜下观察,诱导后细胞核大而圆,核仁明显,胞浆内细胞器发达,可见大量粗面内质网和游离核糖体。提示,MSCs体外可被诱导分化为神经元样细胞,诱导后的细胞有合成某些神经递质的能力并具有发育早期神经元的超微结构特点。     

强迫症含强迫思想和强迫行为者二重辨证施治在开放式心理病房的研究

目的 :探讨二重辨证施治对强迫症含强迫思想和强迫行为者的疗效 ,及其在开放式心理病房的适应情况。 方法:将 5 2例强迫症含强迫思想和强迫行为者按随机的原则在开放式心理病房分为研究组 (2 7例 )和对照组 (2 5例 )。研究组用二重辨证施治 ,对照组用认知疗法结合氯丙咪嗪、氟西汀等。研究时间共 5个月 ,头 2个月为积极治疗期 ,后 3个月为自由治疗期。采用临床疗效评定标准 ,耶鲁布朗强迫症量表 (Y -BOCS) ,x2 检验 ,t检验。 结果 :研究组和对照组 ,积极治疗期末痊愈差异有统计学意义 (x2 =19.94,P <0 .0 0 5 ) ,自由治疗期末痊愈差异有更大的统计学意义 (x2 =2 6.64 ,P <0 .0 0 5 )。耶鲁布朗强迫症量表 (Y -BOCS)总分 ,两组治疗前差异无统计学意义 (t =0 .75 6,P >0 .2 ) ;积极治疗期末 ,两组差异有统计学意义 (t=10 .3 2 4,P <0 .0 0 1) :自由治疗期末 ,两组差异有更大的统计学意义 (t=12 .66,P <0 .0 0 1)。 结论 :二重辨证施治对强迫症含强迫思想和强迫行为者有普遍卓越的痊愈功效和痊愈巩固功效 ,应积极推广。开放式心理病房可作为强迫症含强迫思想和强迫行为者二重辨证施治的医疗平台