携带增强绿色荧光蛋白基因的Id2真核表达载体构建

目的：构建大鼠Id2基因真核荧光表达载体，为骨骼肌的组织工程研究提供有效的分子工具。方法：利用RT-PCR的方法扩增出Id2全长cDNA，利用T4 DNA连接酶将载体pGEM-T和Id2 cDNA进行连接，构建克隆载体，经限制性内切酶EcoR1酶切pGEM-Id2克隆载体和pEGF-C2真核表达载体，构建出重组真核表达载体pEGFP-C2-Id2，经酶切分析、PCR鉴定及DNA测序证实cDNA片段大小和序列的正确性；并通过细胞转染技术将Id2基因导入L6成肌细胞中。结果：经酶切分析和序列测定证实pEGFP-C2-Id2含大小正确的正向Id2 cDNA片段，获取了转染外源性Id2基因的L6细胞。结论：我们成功构建了同时携带有G418筛选位点和增强绿色荧光蛋白的Id2真核表达载体？     

Using Winnicott or Finding Winnicott?

The author suggests, in the title, a difference between using Winnicott's ideas, as in applying them, and finding Winnicott in clinical work. It is in finding examples that resonate to his ideas that we can see most directly what Winnicott had been writing about. And it is at those times of finding Winnicott that his ideas most vividly come alive.     

Kisspeptin levels in idiopathic hypogonadotropic hypogonadism diagnosed male patients and its relation with glucose-insulin dynamic

Male hypogonadism is defined as the deficiency of testosterone or sperm production synthesized by testicles or the deficiency of both. The reasons for hypogonadism may be primary, meaning testicular or secondary, meaning hypothalamohypophyseal. In hypogonadotropic hypogonadism (HH), there is indeficiency in gonadotropic hormones due to hypothalamic or hypophyseal reasons. Gonadotropin-releasing hormone (GnRH) is an important stimulant in releasing follicular stimulant hormone (FSH), mainly luteinizing hormone (LH). GnRH omitted is under the effect of many hormonal or stimulating factors. Kisspeptin is present in many places of the body, mostly in hypothalamic anteroventral periventricular nucleus and arcuate nucleus. Kisspeptin has a suppressor effect on the metastasis of many tumors such as breast cancer and malign melanoma metastases, and is called “metastin” for this reason. Kisspeptin is a strong stimulant of GnRH. In idiopathic hypogonadotropic hypogonadism (IHH) etiology, there is gonadotropic hormone release indeficiency which cannot be clearly described.

A total of 30 male hypogonatropic hypogonadism diagnosed patients over 30 years of age who have applied to Haydarpasa Education Hospital Endocrinology and Metabolic Diseases Service were included in the study. Compared to the control group, the effect of kisspeptin on male patients with hypogonatropic hypogonadism and on insulin resistance developing in hypogonadism patients was investigated in our study.

A statistically significant difference was detected between average kisspeptin measurements of the groups (p?The reason for high kisspeptin levels in the patient group compared to the control group makes us consider that there may be a GPR54 resistance or GnRH neuronal transfer pathway defect. When patients and control groups were compared for HOMA-IR, the difference was not statistically significant. It is considered that kisspeptin is one of the reasons for hypogonatropic hypogonadism and has less effect on insulin resistance.     

急性白血病Id4基因启动子区甲基化状态研究

本研究探讨正常人和不同疾病状态急性白血病（AL）患者Id4基因启动子区甲基化状态差异。采用甲基化特异性聚合酶链反应（MS—PCR）对正常人和AL患者骨髓进行Id4基因启动子区甲基化状况检测。结果表明：Id4基因在正常骨髓中呈完全性非甲基化状态，初治AML和ALL患者中Id4基因甲基化比例分别为84％和86％。8例复发难治的AL患者中Id4基因全部甲基化，14例Id4基因甲基化的完全缓解ALL患者中有8例在12个月内复发，而9例Id4基因非甲基化的完全缓解ALL患者中12月内复发仅1例；Id4基因呈甲基化状态的完全缓解的AML患者中12个月内复发率为62．5％，而在非甲基化的患者中12个月内复发率仅为10％，两者差异有显著性意义。完全缓解的ALL患者甲基化比例为64．3％，而完全缓解的AML患者甲基化比例为28．6％，两者差异有显著性意义。39例初治AL中。有33例Id4基因呈甲基化状态，8例复发难治的AL患者的Id4基因均呈甲基化状态，58例完全缓解的AL中。有24例Id4基因呈甲基化状态。结论：与正常人不同，AL患者中Id4基因都发生了不同程度的甲基化改变，缓解状态患者的甲基化比例低于非缓解状态患者，Id4基因甲基化模式的改变与AL的发生密切相关。     

分化抑制因子Id3对雌性生育能力的研究

目的：通过构建ID3-Gdf9-Cre+的条件性敲除小鼠对雌性小鼠的表型进行研究,从而进一步确定分化抑制因子Id3对雌性生育能力的影响。方法：我们利用配繁出的ID3-Gdf9-Cre+的条件性敲除小鼠,通过HE染色、卵泡计数条件性敲除小鼠卵巢中的卵泡发育的情况进行检测,通过卵母细胞体外成熟实验和体外超排实验对卵母细胞的体外成熟进行检测,通过交配实验对生育力进行检测。结果：通过RT-pcr、WB的结果显示出我们成功在卵母细胞中对ID3进行了敲除;对3W、8W小鼠卵巢进行HE染色并对各级卵泡进行计数,结果表明条敲小鼠中各级卵泡发育正常;通过卵母细胞体外成熟以及体外超排实验显示了条敲小鼠中卵母细胞体外成熟正常;结论：在正常饲养条件下,条件性敲除ID3不影响正常卵泡的发育以及雌性小鼠的生育力。     

银杏内酯B对神经干细胞分化的影响及其机制研究

目的观察银杏内酯B对体外培养的神经干细胞（NSCs）分化的影响。方法用含不同浓度银杏内酯B（20mg／L、40mg／L、60mg／L）的分化培养基培养NSCs 3d和7d，测量细胞突起长度和胞体面积，用免疫荧光检测神经丝蛋白-200（NF-200）、胶质纤维酸性蛋白（GFAP）及少突胶质细胞特异性蛋白（CC-1）、细胞因子信号抑制因子2（SOCS2）、DNA结合抑制因子2（Id2）的表达。结果银杏内酯B组细胞突起增长，胞体增大；NF阳性神经元样细胞百分率增加，GFAP阳性星形胶质样细胞百分率随银杏内酯B浓度增加而增加；SOCS2阳性细胞百分率（35．46％）较对照组（22．17％）明显增加（P〈0．01），银杏内酯B组Id2阳性细胞百分率（51．52％）较对照组（66．24％）明显减少（P〈0．01）。结论银杏内酯B可促进分化细胞成熟，促进NSCs向神经元分化；星形胶质细胞百分率的增高与银杏内酯B呈剂量依赖关系。