p38MAPK在17β-雌二醇和雷洛昔芬促成骨细胞増殖和分化中的作用

目的:观察p38MAPK信号转导通路在17β-雌二醇和雷洛昔芬促成骨细胞增殖和分化过程中的作用。方法:取第一继代BALB/c小鼠头盖骨成骨细胞,药物刺激组分别加入不同浓度17β-雌二醇或雷洛昔芬;含阻断剂组预先添加不同浓度的SB202190(p38MAPK的阻断剂)阻断信号转导通路,再加17β-雌二醇或雷洛昔芬,作用72h后用MTT法与PNPP法测定细胞的增殖能力和碱性磷酸酶活性。结果:加入雌激素或雷洛昔芬后,成骨细胞的增殖和分化明显增强,与空白对照组比较差异具有显著性意义(P<0·05);阻断p38MAPK信号转导通路后,细胞增殖分化受到明显抑制,与未阻断组比较差异具有非常显著性意义(P<0.01)。结论:p38MAPK在17β-雌二醇和雷洛昔芬诱导的小鼠成骨细胞的増殖和分化过程中发挥重要作用。     

体外诱导小鼠骨髓单个核细胞向内皮分化的实验研究

目的:为组织工程研究做准备,分离培养小鼠骨髓单个核细胞并诱导向血管内皮分化。方法:成年Balb/c小鼠,应用淋巴细胞分离液(密度1.077g/m l),将长骨骨髓经F icoll非连续密度梯度离心,收集中层的单个核细胞,加入诱导培养基并置于纤维连接素包被的培养板上进行诱导分化,观察细胞生长状况,以透射电镜及Ⅷ因子、CD31、Lectin免疫组化以及流式细胞仪方法对培养细胞进行鉴定。结果:细胞圆形、纺锤形单层融合贴壁生长;Ⅷ因子、CD31、Lectin免疫组化染色阳性;透射电镜显示细胞具有内皮特征性的W-P小体。结论:采用F icoll密度梯度离心可获得较高纯度的骨髓单个核细胞,经体外培养并诱导分化,具有血管内皮细胞的特征。     

大鼠NSCs向多巴胺能神经元分化研究

目的：探讨IL-1β对神经干细胞（NSCs）分化为多巴胺能神经元作用：方法：体外培养和鉴定中脑来源NSCs，用免疫学方法检测分化细胞酪氨酸羟化酶（TH）表达.结果：血清组、IL-1β10pg／ml组、IL-1β120pg／ml组、IL-1β150pg／ml组、IL-1β200pg／ml组诱导7d后TH细胞分别平均为0,45％、0．6％、7.2％、12．5％、8．75％：结果显示IL-1β诱导NSCs明显提高TH细胞表达率，与血清组比较有显著性差异（P〈0，05），在IL-1β150pg／ml剂量范同内TH细胞表达率与剂量呈正相关。结论：IL-1β有诱导NSC向多巴胺能神经元分化的显著作用：     

湿温汗下润忌宜及儿科临床

<温病条辨>上焦篇湿温门云:"头痛恶寒,身重疼痛,舌白不渴,脉弦细而濡,面色淡黄,胸闷不饥,午后身热,状若阴虚,病难速已,名日湿温,汗之则神昏耳聋,甚则目瞑不欲言,下之则洞泄,润之则病深不解".     

非小细胞肺癌中伴神经内分泌分化及c-erbB-2表达的意义

 目的　探讨非小细胞肺癌(NSCLC) 伴神经内分泌分化(NE) 与原癌基因c-erbB-2 表达的关系。方法　观察非小细胞肺癌32 例,用免疫组织化学S-P 法来判定神经内分泌分化及c-erbB-2 表达特点。结果　NSCLC 中24 例有NE 分化,c-erbB-2 在伴NE 分化和无NE 分化的表达阳性率比较,差异有显著性,NE 分化的阳性率和癌细胞分化程度有显著性差异。c-erbB-2 阳性在淋巴结转移组表达与无淋巴结转移组表达相比有显著性差异。结论　c-erbB-2 的过表达在非小细胞肺癌中与其侵袭性的生物学行为密切相关。c-erbB-2 过表达可用来判断NE 分化的非小细胞肺癌预后。     

男性原发性骨质疏松症中医证型的实质探讨

目的:探讨男性原发性骨质疏松症中医证型的实质.方法:男性原发性骨质疏松症患者按中医临床辩证分型为肾阳虚型、肾阴虚型和非肾虚型.采用回归和判别分析法来研究骨密度、血清激素、血生化、血清微量元素、尿生化等指标与中医证型的相关性,探讨其中医证型的实质.结果:男性原发性骨质疏松症患者睾酮(T)、血清降钙素(CT)、铜(Cu)值显著降低;肾阴虚组、肾阳虚组骨钙素(BGP)值均显著高于对照组和非肾虚组;CT值于各组之间差别显著,其中非肾虚组CT值显著低于正常对照组,肾阴虚组、肾阳虚组CT值均非常显著低于对照组;甲状旁腺激素的羧基端片段(PTH-c)值于各组之间差别显著,肾阴虚组、肾阳虚组PTH-c值均非常显著低于对照组和非肾虚组,其中肾阳虚组还显著低于肾阴虚组;雌二醇(E2)值于各组差别显著,其中非肾虚组显著低于正常对照组,肾阴虚组、肾阳虚组非常显著低于对照组.结论:男性原发性骨质疏松症患者钙丢失、骨丢失明显加快,而骨形成并没有加快.肾虚与非肾虚划分的生化基础为T和尿钙/肌酐(Ca/Cr);男性原发性骨质疏松症的中医证型与骨代谢生化指标有密切相关,骨代谢生化指标的改变是辩证分型的基础.     

中医学发展有赖于对病证关系的再认识

提出就中医学辨证与辨病的关系,应该是方法与结果的关系.辨证必须以辨病为前提,证只有因病的存在才有自己的特殊性可言.而这时的病不能是传统意义上的以症状命名的"病",而必须是具有发展规律、表现出特定症状与相应阶段证候的"新"病.     

何世东治疗强直性脊柱炎临床经验

强直性脊柱炎是脊柱关节炎常见的临床类型,好发于青年男性,以中轴关节受累为主,可伴发关节外表现,严重者可发生脊柱畸形和关节强直,是一种慢性炎症性疾病。何老临证潜心研究,注重强直性脊柱炎病变的核心,从临床常见的证型着手,攻克难治之疾,何老认为该病根源在于肾虚,肾虚累及肝脾,脾胃不足则后天之气血精微无以生化,筋脉骨骼失去滋养。肝不足则血虚,继而筋脉失养,发生该病。     

Purmorphamine increased adhesion,proliferation and expression of osteoblast phenotype markers of human dental pulp stem cells cultured on beta-tricalcium phosphate

ObjectivesGrowth factors play a significant role in cell proliferation and differentiation during different stages of the bone repair. However, several limitations have been brought researchers attention to an osteoinductive small molecule including Purmorphamine. In this study, we aimed to evaluate the effect of Purmorphamine on adhesion, proliferation and differentiation of human dental pulp stem cells (hDPSCs) seaded on beta-tricalcium phosphate (β-TCP) granules.MethodshDPSCs were established from extracted wisdom teeth of healthy volenteers. Cells at passage 3 were seeded on β-TCP in the presence or absence of Purmorphamine. Cell adhesion and proliferation were assessed using scanning electeron microscopy (SEM) and DNA counting assay, respectively, after 1, 3 and 5 days. Then, hDPSCs seeded on β-TCP were subjected to osteogenic medium with or without Purmorphamine. After 7 and 14 days osteogenic diffrentiation capability of hDPSCs were determined using real-time RT-PCR and alkaline phosphatase (ALP) activity assay.ResultsThe significant increase in amount of DNA was observed at day 3 and 5 in the presence of Purmorphamine. SEM imaging also was confirmed the DNA counting assay; in all given time points, hDPSC attachment and growth was significantly higher in the presence of Purmorphamine. ALP activity was increased by Purmorphamine at both 7 and 14 days of induction. Purmorphamine showed to effect on osteopontin expression at earlier stage of osteogenic differentiation, whereas for osteocalcin expression, this effect was more evident at later stage of differentiation.ConclusionPurmorphamine had a promotive effect on adhesion, proliferation and osteogenic differentiation of hDPSCs cultured on β-TCP. The outcome of the current study would help in development of in vitro culture conditions for better osteogenic differentiation of hDPSCs prior to transplantation.