High-plasticity mesenchymal stem cells isolated from adult-retained primary teeth and autogenous adult tooth pulp—A potential source for regenerative therapies?

PurposeThe objective of this study was to compare the growth rate, morphology, immunohistology and plasticity of autogenous adult-retained SHEDs (arSHEDs) and adult dental pulp stem cells (DPSCs) obtained from the same donor.MethodsExpression of the mesenchymal stem cell markers CD44, CD90, CD105, caspase-3 and GAPDH were assessed using RT-PCR. Caspase-3 and CD44 were also evaluated at the protein level by western blotting of cell lysates. Plasticity of DPSCs and arSHEDs were tested by culture in adipogenic, chondrogenic, osteogenic and Schwann cells induction media.ResultsDPSCs and arSHEDs were isolated by explant culturing and were similarly positive for growth rate and all tested markers. Furthermore, DPSCs and arSHEDs could be driven to adipocyte, chondrocyte, osteocyte and Schwann cells lineages thus indicating similar plasticity as precursor cells.ConclusionThis study demonstrates the similarities between DPSCs and arSHEDs in a unique situation, where both stem cells (SC) types were obtained from a single patient and thus represent an alternative source of SC’s for tissue engineering and regeneration.     

The effects of graphene nanostructures on mesenchymal stem cells

We report the effects of two-dimensional graphene nanostructures; graphene nano-onions (GNOs), graphene oxide nanoribbons (GONRs), and graphene oxide nanoplatelets (GONPs) on viability, and differentiation of human mesenchymal stem cells (MSCs). Cytotoxicity of GNOs, GONRs, and GONPs dispersed in distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)] (DSPE-PEG), on adipose derived mesenchymal stem cells (adMSCs), and bone marrow-derived mesenchymal stem cells (bmMSCs) was assessed by AlamarBlue and Calcein AM viability assays at concentrations ranging from 5 to 300 μg/ml for 24 or 72 h. Cytotoxicity of the 2D graphene nanostructures was found to be dose dependent, not time dependent, with concentrations less than 50 μg/ml showing no significant differences compared to untreated controls. Differentiation potential of adMSCs to adipocytes and osteoblasts, – characterized by Oil Red O staining and elution, alkaline phosphatase activity, calcium matrix deposition and Alizarin Red S staining – did not change significantly when treated with the three graphene nanoparticles at a low (10 μg/ml) and high (50 μg/ml) concentration for 24 h. Transmission electron microscopy (TEM) and confocal Raman spectroscopy indicated cellular uptake of only GNOs and GONPs. The results lay the foundation for the use of these nanoparticles at potentially safe doses as ex vivo labels for MSC-based imaging and therapy.     

中医论治血管性头痛

随着生活节奏的加快、生活压力的不断加大，以临床多发、病因繁杂、反复难愈为特征的血管性头痛，给工作、学习造成困扰不断增大。现代医学多对症治之，并无特效根治之法。近年来，随着以完备的中医基础理论为指导的传统医学蓬勃发展，逐渐形成以头痛性质、部位为主导的症状论治，以血瘀、痰饮、六淫邪气、七情内伤为主导的致病因素论治和以肝、脾、肾为主导的脏腑论治的基本构架，由外而内构建起了以“症状-致病因素-脏腑”为核心的三维立体论治策略，对临床具有指导意义。     

A new rosette in retinoblastoma

Retinoblastoma, the most common primary malignant intraocular tumor of childhood is a great success story in pediatric and ocular oncology. Pathology of retinoblastoma is important to guide the treatment modalities. Differentiated retinoblastoma is commonly seen in younger age group. Since a hundred years, we have been observing two typical true rosettes in retinoblastoma in the form of Flexner-Wintersteiner (FW) and Homer Wright (HW) rosettes and in many occasions pseudorosettes have been documented. In the present case report, a third new type of rosette was identified in a differentiated retinoblastoma which had an unusual anterior segment involvement.     

唾液腺恶性多形性腺瘤HER2基因扩增与临床病理特征及预后的关系

目的: 检测唾液腺恶性多形性腺瘤（malignant pleomorphic adenoma,MPA）中人上皮生长因子受体2（human epithelial growth factor receptor 2,HER2）蛋白表达、基因扩增情况,分析其与肿瘤临床病理及患者预后的相关性,以期评估其作为靶向治疗位点的可能性。方法: 应用免疫组织化学染色法检测140例MPA患者肿瘤组织中HER2蛋白的表达,对HER2基因扩增情况不明确者（HER2 2+）行荧光原位杂交,确定基因扩增状态。统计140例患者临床病理资料并进行随访,应用SPSS16.0软件包中的Kaplan-Meier、Cox比例风险模型,分析HER2基因扩增与MPA肿瘤临床病理及患者预后的相关性。结果: 140例MPA患者中,HER2阳性率为25%。腺癌亚型MPA中,HER2阳性率达到40.5%（32/79）,显著高于肌上皮癌亚型MPA（4.9%,3/61）（P<0.05）。HER2阳性与肿瘤患者性别、组织学分级及N分期显著相关。生存分析显示,HER2阳性患者总生存率及疾病别生存率低。结论: HER2基因过表达及扩增与MPA肿瘤细胞恶变亚型显著相关,并且提示MPA患者较差的预后状态。     

维甲酸、地塞米松和生长激素释放激素联合诱导生长激素细胞分化

目的探讨维甲酸(RA)、地塞米松(Dex)和生长激素释放激素(GHRH)在诱导生长激素(GH)细胞分化中的作用。方法原代培养大鼠胚胎垂体细胞,将RA(10-6mol/L)、Dex(50 nmol/L)和GHRH(10-7mol/L)单独或联合应用6 d,诱导大鼠胚胎垂体细胞分化,利用免疫组化检测GH阳性细胞百分比,放射免疫分析检测培养液的上清液中GH含量。结果与对照组相比,单独应用RA显著增加GH阳性细胞的百分比和GH的含量(P<0.05),而单独应用Dex和GHRH均无此作用(P>0.05);RA诱导4 d后,再加入Dex诱导2 d,其诱导效应较单独应用RA明显增强(P<0.05),而加入GHRH则不起作用(P>0.05);RA和Dex联合诱导4 d后,再加入GHRH诱导2 d,其诱导效应较RA和Dex联合诱导明显增强(P<0.01)。结论RA能够促进GH细胞的早期分化,Dex和GHRH则不能;RA能够与Dex、GHRH发挥协同效应,诱导GH细胞分化。     

我对韩国火病(Hwa-Byung)的认识

火病(Hwa-Byung)是韩国人很熟悉的心身疾病之一,属于韩国人固有文化相关综合征。反而此韩国火病在中医界并不熟知,本文收集和整理了目前有关火病的内容,介绍韩国火病的实际现状。韩国庆熙医疗中心韩方神经精神科通过使用choseunghun开发软件即"火病认定的标准诊断协议",在2004年5月至11月以门诊患者为主进行研究,确定患者辨火病的辨证类型结果如下,其中最多的辨证类型心阴阳两虚证(15),气血两虚证(5),少阳邪气弥漫三焦证(8),热扰胸膈证(8),痰火扰心证(7)。     

EFFECT OF GROWTH FACTORS ON DNA LABELING AND CYTOSKELETAL PROTEIN EXPRESSION IN 17-β-ESTRADIOL AND BASIC FIBROBLAST GROWTH FACTOR PRE-TREATED ASTROCYTE CULTURES

Astrocytes react to all noxae which damage neurons. Their reactions include degeneration, hypertrophy, hyperplasia and fibre formation. Growth factors inducing proliferation and differentiation of both neurons and astrocytes in culture play a pivotal role in the dynamic flow of signaling molecules between neurons and astroglia. Estrogens as well influence astroglia and are neuroprotectants. This study has investigated the interactions between growth factors and estrogens on DNA labeling and cytoskeletal protein [glial fibrillary acidic protein (GFAP) and vimentin] expression in 22 DIV astrocyte cultures treated for 24 or 36?h under different experimental conditions.

Contemporary addition of 17-β-estradiol (E₂) with two or three growth factors for 24?h, significantly stimulated methyl-[³H]thymidine incorporation into DNA from 22 days in vitro (DIV) astrocyte cultures.

This effect reached a peak when E₂ was co-added with epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and insulin. In astrocyte cultures treated for 36?h with E₂ and EGF+insulin or bFGF+insulin added in the last 12?h, DNA labeling was remarkably increased. The parallel cyclin D1 expression positively correlated with ERK2 activation. Western blot analysis for cytoskeletal proteins showed also changes of both GFAP and vimentin expression. The above data suggest the occurrence of a scheduled interaction between “competence” or “progression” growth factors and estrogens on DNA labeling and cytoskeletal protein expression during astroglial cell proliferation and differentiation in culture. A better understanding of the mechanisms of these interactions may contribute to develop strategies for controlling astroglial reaction in cerebrovascular disease including stroke and hypertensive brain damage.     

氨基甾体H42649对K562白血病细胞的抑制增殖和诱导分化作用

目的观察氨基甾体H42649对人慢性粒细胞白血病K562细胞系的抑制增殖和诱导分化作用。方法采用液体培养实验,MTT实验,集落培养实验观察10-8～10-4mol/L浓度的H42649对K562细胞增殖能力的影响;采用Wright-Giemsa染色,联苯胺染色和流式细胞术评价10-6mol/L浓度的H42649对K562细胞的诱导分化作用。结果不同浓度的H42649连续作用K562细胞1～5 d后,细胞计数和集落计数明显减少;第5 d处理组的MTT值显著低于对照组,并呈剂量依赖关系;10-6mol/L浓度的H42649对K562细胞作用5d后,联苯胺染色A值升高(P<0.01),形态学观察其趋向成熟分化,流式细胞术检测药物处理4 d后K562细胞膜上CD71表面标记表达阳性率为84%。结论氨基甾体H42649能显著抑制K562细胞的增殖并诱导其向红系分化,提示该药可作为一种新型慢粒白血病细胞的诱导分化剂。     

Skeletal muscle growth defect in human growth hormone transgenic rat is accompanied by phenotypic changes in progenitor cells

Growth hormone (GH) is known to have a pivotal role in the maintenance of skeletal muscle mass. Sarcopenia, the loss of skeletal muscle mass, is a common phenomenon in aging, and it is widely accepted that sarcopenia is largely attributed to age-related decline in GH secretion. In the present study, we tested if human growth hormone transgenic rats (GH-TG rats) whose plasma GH levels are maintained relatively low could be an appropriate model for sarcopenia. Analyses of GH-TG rats revealed that they exhibit skeletal muscle growth defect as well as atrophy of myofibers. The number of myofibers in tibialis anterior muscle was comparable to that of WT rats, while the proportion of type I slow myofibers in tibialis anterior muscle was increased in GH-TG rats after 5 months. Neither increased expression of ubiquitin ligases, MuRF1 and MAFbx, nor indication of apoptotic cell death was observed. Notably, myogenic differentiation potential of skeletal muscle progenitor cells in GH-TG rats was lower than WT rats, and this was accompanied by increased adipogenic potential. These results indicate that GH-TG rats could be a useful model to elucidate the mechanism of sarcopenia induced by reduced GH action and raised the possibility that decreased GH action may cause an alteration of differentiation potential of skeletal muscle progenitor cells.