Bilateral isolated Epididymal Agenesis in a 32 year old man

Epididymal agenesis is defined as the absence of the epididymis totally or segmentally, unilateral or bilateral, which is secondary to the Wolffian duct malformation (1). Rete testis, epididymis, vas deferens and seminal vesicle are believed to develop from Wolffian ducts.     

尿路上皮细胞和膀胱脱细胞基质的相容性研究

目的 研究尿路上皮细胞(UCs)和膀胱脱细胞基质(BAM)的生物相容性.方法 分离培养兔UCs,以1.0×105/cm2均匀接种于BAM上,观察细胞的粘附、生长及增殖.将复合物种植于兔背部皮下检测其组织相容性.结果 典型的UCs呈多角形的“铺路石样”结构,抗CK AE1/AE3抗体免疫组化阳性证实为上皮细胞;BAM内部未见细胞碎片,可见排列规则而紧密连接成网状的胶原纤维;细胞种植到支架上一周后,行HE染色检测显示膀胱上皮细胞在BAM上贴附生长良好;生长曲线显示,实验组和对照组生长曲线相似.结论 UCs在BAM表面生长良好,相容性较好,是组织工程良好的种子细胞.     

应用高效液相色谱串联质谱法检测细胞内神经酰胺的含量

目的采用高效液相色谱串联质谱法建立测定细胞中不同类型神经酰胺含量的分析方法.方法采用Agilent 1290高效液相色谱仪系统, ZORBAX Eclipse XDB-C8(150.0 mm×2.1 mm, 3.5 μL)色谱柱, 柱温35 ℃; 流动相A为含1 mmol/L甲酸铵和0.2%甲酸的甲醇溶液, 流动相B为100%甲醇; 流速为0.3 mL/min, 梯度洗脱; 质谱检测中采用电喷雾离子源多反应监测模式.结果C16、C17、C18、C24四种神经酰胺的色谱响应线性范围良好( R ² ≥0.9989);平均回收率分别为99.1%、99.9%、100.5%、98.2%, 相对标准偏差分别为5.6%、5.1%、4.8%、5.5%( n = 9). 结论采用高效液相色谱串联质谱法建立同时测定细胞中不同类型神经酰胺的方法灵敏度高, 重现性好, 可以作为检测细胞中不同类型神经酰胺含量的工具.     

脂肪干细胞对大鼠胰岛细胞脂性凋亡的保护作用

[目的]探讨脂肪干细胞(ADSCs)对棕榈酸(PA)诱导大鼠胰岛瘤细胞(INS-1)凋亡的保护作用.[方法]本实验分为三组.空白对照组:正常培养液+INS-1单独培养(其上层只含培养基);PA组:含0.5 mmol/L PA的高脂培养液+INS-1单独培养;INS-1+ ADSCs组:高脂培养液+INS-1+ ADSCs共培养.各组在处理24 h后检测各组细胞的存活率、胰岛细胞基础胰岛素分泌量(BIS)和葡萄糖刺激的胰岛素分泌量(GSIS);采用RT-PCR检测胰岛细胞中Ins-1、Glut-2、Pdx-1表达量;采用Elisa检测各组IL-6、TNF-α、HGF、TIMP-1和TGF-β水平.[结果]PA组和PA+ ADSCs组细胞存活率均较对照组下降(P＜0.01),但PA+ ADSCs组细胞存活率明显大于PA组(P＜0.01).与对照组相比,PA组和PA+ ADSCs组BIS增加(P＜0.01),而GSIS均减少(P＜0.01);与PA组比较,PA+ ADSCs组细胞BIS差异无统计学意义(P＞0.05),而GSIS显著增加(P＜0.01).和对照组相比,PA组和PA+ ADSCs组的Pdx-1、Ins-1、Glut-2 mRNA水平均明显下降(P＜0.01);和PA相比,PA+ ADSCs组细胞的Pdx-1和Glut-2 mRNA水平明显上升(P＜0.01),而Ins-1 mRNA水平比较差异无统计学意义(P＞0.05).PA+ ADSCs组中IL-6、TNF-α、HGH、TIMP-1、TGF-β的含量均较其他两组明显升高.[结论]PA能诱导INS-1细胞凋亡并抑制Ins-1、Glut-2、Pdx-1的表达,而ADSCs可以通过旁分泌机制并上调Glut-2和Pdx-1的表达对胰岛细胞起到保护作用.     

人子宫内膜细胞体外培养模型的建立

【目的】构建人子宫内膜细胞体外培养模型。【方法】收集子宫内膜组织,采用酶消化、筛网滤过、梯度离心的方法获得腺上皮与基质细胞混合细胞进行体外原代培养,采用光学显微镜下观察体外培养的细胞形态,采用波形蛋白和角蛋白单克隆抗体进行免疫组化法鉴定子宫内膜基质细胞和腺上皮细胞。【结果】倒置显微镜下发现子宫内膜细胞中腺上皮细胞和基质细胞呈混合生长,其中基质细胞占大部分,为72．0％;腺上皮细胞占小部分,为24．5％,基质细胞和腺上皮细胞免疫组化染色后呈阳性,细胞纯度分别为24．5％和70．5％,两种细胞加起来的纯度为95％。【结论】该方法操作简便,效率高,可建立一个稳定实用的人子宫内膜细胞体外培养体系。     

干细胞相关因子在大鼠卵巢颗粒细胞中的表达

背景：现有研究表明,人与大鼠卵巢优势卵泡中的颗粒细胞具有表达干细胞特性的现象。目的：探讨干细胞相关因子在大鼠卵巢颗粒细胞中的表达。方法：将大鼠卵巢组织制作石蜡切片后,使用免疫组化法检测CD34,CD133,ABCG2/Bcrp1,Pou5f1/Oct-4的表达。卵泡穿刺法获取颗粒细胞,并使用免疫组化法检测FSHR受体的表达以鉴定颗粒细胞的纯度。免疫组化法检测颗粒细胞 CD44, C-Kit 的表达情况, RT-PCR 检测卵巢组织与颗粒细胞 ABCG2/Bcrp1、Pou5f1/Oct-4、Nanog基因的表达情况。结果与结论：免疫组化法石蜡切片显示部分卵巢颗粒细胞CD34,CD133,ABCG2/Bcrp1,Pou5f1/Oct-4阳性表达,而Pou5f1/Oct-4蛋白的表达在卵泡发育的周期中逐步增多,并在黄体化的时候显著增强,形成白体后则消失,因而具有周期性表达的特点。卵泡穿刺法提取的原代颗粒细胞FSHR受体鉴定阳性率在95%以上。体外培养的颗粒细胞以长梭形或菱形为主,部分细胞有聚集生长现象,且 CD44,C-Kit 阳性表达。RT-PCR检测结果显示,卵巢组织与体外培养的颗粒细胞均不表达 Nanog,卵巢低表达 Pou5f1/Oct-4,强烈表达ABCG2/Bcrp1,颗粒细胞微弱表达Pou5f1/Oct-4,而ABCG2/Bcrp1表达较强。以上结果显示大鼠卵巢中的部分颗粒细胞具有干细胞的特性。     

The Regenerative Medicine in Oral and Maxillofacial Surgery: The Most Important Innovations in the Clinical Application of Mesenchymal Stem Cells

Regenerative medicine is an emerging field of biotechnology that combines various aspects of medicine, cell and molecular biology, materials science and bioengineering in order to regenerate, repair or replace tissues.The oral surgery and maxillofacial surgery have a role in the treatment of traumatic or degenerative diseases that lead to a tissue loss: frequently, to rehabilitate these minuses, you should use techniques that have been improved over time. Since 1990, we started with the use of growth factors and platelet concentrates in oral and maxillofacial surgery; in the following period we start to use biomaterials, as well as several type of scaffolds and autologous tissues. The frontier of regenerative medicine nowadays is represented by the mesenchymal stem cells (MSCs): overcoming the ethical problems thanks to the use of mesenchymal stem cells from adult patient, and with the increasingly sophisticated technology to support their manipulation, MSCs are undoubtedly the future of medicine regenerative and they are showing perspectives unimaginable just a few years ago. Most recent studies are aimed to tissues regeneration using MSCs taken from sites that are even more accessible and rich in stem cells: the oral cavity turned out to be an important source of MSCs with the advantage to be easily accessible to the surgeon, thus avoiding to increase the morbidity of the patient.The future is the regeneration of whole organs or biological systems consisting of many different tissues, starting from an initial stem cell line, perhaps using innovative scaffolds together with the nano-engineering of biological tissues.     

Neural stem cells and alcohol

This article summarizes the proceedings of a symposium held at the 2002 Research Society on Alcoholism Meeting in San Francisco, California. The aim of this symposium was to review research on the effects of ethanol on neural stems cells and neurogenesis. Ethanol is known to alter neurogenesis during development; however, recent studies indicate that the brain forms new neurons from stem cells throughout life. Furthermore, stem cells can be transplanted into the brain, creating exciting new possibilities to study brain function. The symposium covered these research areas. Dr. Michael W. Miller reviewed knowledge on the effects of ethanol on stem cell proliferation and differentiation during development. Dr. Wu Ma described studies in culture indicating that (1) neural stem cells express functional muscarinic acetylcholine receptors (mAchR), (2) mAchR-mediated proliferation involves Ca signaling and mitogen-activated protein kinase phosphorylation, and (3) phosphoinositol-3 kinase is a downstream effector for mAchR-mediated cell proliferation via activation of Akt. Drs. Kim Nixon and Fulton T. Crews followed with in vivo studies on ethanol's effects on adult neural stem cell proliferation and differentiation. Dr. W. Michael Zawada described studies directed at dopamine neuron cell transplants into mammalian central nervous system. These studies clearly establish that ethanol has significant effects on stem cells.     

下肢动脉硬化性闭塞症患者外周血内皮祖细胞数量和功能的变化

目的 研究下肢动脉硬化性闭塞症患者外周血内皮祖细胞数量及其功能的改变.方法 入选对象60例分为实验组(n=30)和对照组(n=30).密度梯度法分离人外周血单个核细胞,诱导分化培养7天后,荧光染色和流式细胞术鉴定贴壁细胞为内皮祖细胞.采用吉姆萨染色法计算细胞个数、集落数目,MTT比色法、改良Boyden小室及黏附能力实验测定内皮祖细胞增殖、迁移和黏附能力.结果 下肢动脉硬化性闭塞症患者外周血内皮祖细胞数量较对照组(27.2±3.6:52.6±5.9,细胞/×200倍视野,P<0.05)减少,集落个数较对照组(16.6±4.8:22.3±4.9,集落/×40倍视野,P<0.05)也减少,而且增殖能力 (0.193±0.064:0.243±0.078,P<0.05)、迁移能力(12.1±2.7:17.8±4.2,细胞/×200倍视野,P<0.05)、黏附能力(47.3±4.3:51.9±3.7,细胞/×200倍视野,P<0.05)受损.结论 下肢动脉硬化性闭塞症患者外周血内皮祖细胞数量减少,生物学功能减退.