流行性出血热患者骨髓细胞超微结构的研究

8例流行性出血热患者骨髓细胞的超微结构研究结果发现骨髓细胞出现变性、空泡、粗面内质网及高尔基体扩张。并在胞浆中发现颗粒-丝状病毒包涵体及直径为100nm的病毒颗粒。同时胶体金免疫电镜发现骨髓细胞EHF抗原阳性,说明骨髓细胞是其繁殖、复制的场所。     

Evidence for cell-specific changes with age in expression of oestrogen receptor (ER) alpha and beta in bone fractures from men and women

Oestrogen is recognized as important for maintaining bone mass in men and women. Oestrogen receptor (ER) alpha and the recently described ER-beta are both expressed in bone cells, but have different affinities for oestrogen agonists and plant oestrogens, which could be important in developing treatments for bone loss in both men and women. It is unclear, however, which isoform predominates in bone; cell type and age may influence their relative expression. The present study has compared ER-alpha and ER-beta expression in serial sections of human fracture callus from males (n = 19, age range 5-72 years) and females (n = 15, age range 3-86 years) by indirect immunoperoxidase. Fracture callus was used as it can be readily obtained from individuals over a wide age range and contains a variety of bone cells. Antibody specificity was confirmed by western blotting and comparison of immunoreactivity in sections of breast tumour and benign prostate hyperplasia. No gender difference in ER expression was found in bone from individuals less than 40 years old. Proliferative chondrocytes were positive for both isoforms, but few larger hypertrophic cells were immunoreactive. ER-alpha and ER-beta were co-expressed in osteoclasts, suggesting that oestrogen may act directly on these cells. Osteoblasts, osteocytes, and mesenchymal cells also expressed both isoforms. In women over 40 years of age, however, relatively fewer biopsies contained osteocytes positive for ER-alpha and ER-beta. Likewise, the proportions of osteoblasts and mesenchymal cells expressing ER-beta were reduced but ER-alpha remained unaffected. In contrast, in men over 40 years, only the proportion of biopsies containing ER-beta-positive mesenchymal cells was lower. In these older men and women, ER-alpha and ER-beta expression was retained by the small proliferative chondrocytes. These results demonstrate that gender, age, and cell type are important determinants of ER isoform expression in skeletal cells.     

鼻腔、鼻窦部砂粒性骨化纤维瘤6例临床分析

为探讨临床上比较罕见的砂粒性骨化纤维瘤（psammomatoid ossifying fibroma,POF）的临床特征、诊断与治疗方法,分析了6例POF的临床资料并综合文献,提出POF是一种以青少年为主、具有局部侵袭性的良性肿瘤,有明确的组织形态特征,临床症状特别是CT扫描是临床诊断的基础,治疗原则是在彻底清除病灶的基础上,兼顾面容与功能的保护。     

普拉固对兔激素性股骨头坏死骨细胞凋亡的干预研究

目的:采用激素(地塞米松,Dexamethasone)制作成兔股骨头坏死(Steroid-induced Femoral Head Necrosis, SIFHN) 模型,探讨普拉固(Pravastatin)是否能干预SIFHN中骨细胞凋亡.方法:12～18周龄日本大白兔34只,雌雄不限,体重(2.5±0.6) kg,随机分为3组:对照组(A组):6只,单纯标准饲料及草料喂养.实验组(B组):14只,Dexamethasone 2.5 mg/kg·d-1 肌注.干预组(C组):14只,Dexamethasone 2.5 mg/kg·d-1肌注 普拉固2.5 mg/kg·d-1口服.期间每组均予青霉素80万单位/只,肌注,链霉素1.0 g/只,肌注,2次/周,防止感染.分别于6、8、10、12周处死每组兔子各3只,实验开始及处死前均检查血脂、血Prothrombin Time(PT)、tissue plasminogen(t-PA),处死后行光镜下细胞凋亡分析.结果:C组的兔血脂较B均降低(P<005),PT较实验组均降低 (P<0.05),t-PA较B组升高(P<0.01).与B组比较,光镜示细胞凋亡分析指数减少(P<0.01).结论:地塞米松能在兔体内成功造成股骨头坏死模型,而他汀类药物普拉固能有效干预SIFHN中骨细胞凋亡,其作用机制可能与降脂、抗凝、调控凋亡基因表达等因素有关.     

Piezo1离子通道介导的流体剪切应力抑制MLO-Y4骨细胞凋亡

目的:研究Piezo1机械敏感性离子通道对骨细胞的调控作用。方法:应用课题组研发的流体剪切力（FSS）加载装置,使用免疫荧光探究FSS对Piezo1通道在MLO-Y4中影响的最佳强度;对Piezo1使用FSS、激动剂Yoda1或阻滞剂GsMTx4进行干预。使用Western Blot探究Cleaved Caspase-3、Bcl-2、Bax凋亡相关蛋白的表达量;使用Hoechst-33258以及流式细胞凋亡检测MLO-Y4骨细胞的凋亡比例。结果:免疫荧光染色结果表明FSS对MLO-Y4骨细胞中Piezo1的表达呈现先增加后降低的单峰趋势,其中以9 dyne/cm2加载30 min为最佳刺激条件。使用Yoda1抑制MLO-Y4细胞凋亡（P<0.05）,使用GsMTx4促进MLO-Y4细胞凋亡（P<0.05）。Piezo1介导的FSS抑制MLO-Y4细胞凋亡（P<0.05）。结论:Piezo1机械敏感性离子通道介导的FSS在9 dyne/cm2条件下抑制MLO-Y4骨细胞凋亡,且FSS在该条件下可以有效逆转Piezo1阻滞剂GsMTx4在4 μmol/L作用0.5 h下促进MLO-Y4细胞凋亡作用。     

构建及鉴定人生存素基因的慢病毒表达载体

背景：抑制椎间盘细胞的凋亡可以延缓椎间盘的退变,而生存素具有调节细胞增殖和抗凋亡功能。目的：构建人生存素基因的慢病毒载体。方法：应用全基因合成技术合成人生存素基因（BIRC5）,通过 PCR扩增目的基因并对 PCR 结果进行电泳分析。将目的基因克隆到慢病毒表达质粒构建重组慢病毒质粒 Lenti-BIRC5。将重组的慢病毒质粒转化细菌感受态细胞,PCR鉴定阳性的克隆进行基因测序。将含有目的基因的慢病毒质粒转染293T细胞,应用Western blot技术对重组慢病毒载体 Flag-Survivin 融合蛋白的表达进行检测。结果与结论：PCR鉴定及基因测序结果显示成功构建了含有人Survivin基因的慢病毒表达载体,Western blot检测结果显示目的基因在体外培养细胞中转染成功并且过表达。说明慢病毒表达载体Lenti-BIRC5构建成功,这为下一步研究Survivin在人髓核细胞中的抗凋亡作用提供了载体。     

Parathyroid hormone (PTH)–induced bone gain is blunted in SOST overexpressing and deficient mice

Intermittent parathyroid hormone (PTH) treatment is a potent bone anabolic principle that suppresses expression of the bone formation inhibitor Sost. We addressed the relevance of Sost suppression for PTH‐induced bone anabolism in vivo using mice with altered Sost gene dosage. Six‐month‐old Sost overexpressing and 2‐month‐old Sost deficient male mice and their wild‐type littermates were subjected to daily injections of 100 µg/kg PTH(1–34) or vehicle for a 2‐month period. A follow‐up study was performed in Sost deficient mice using 40 and 80 µg/kg PTH(1–34). Animals were sacrificed 4 hours after the final PTH administration and Sost expression in long bone diaphyses was determined by qPCR. Bone changes were analyzed in vivo in the distal femur metaphysis by pQCT and ex vivo in the tibia and lumbar spine by DXA. Detailed ex vivo analyses of the femur were performed by pQCT, µCT, and histomorphometry. Overexpression of Sost resulted in osteopenia and Sost deletion in high bone mass. As shown before, PTH suppressed Sost in wild‐type mice. PTH treatment induced substantial increases in bone mineral density, content, and cortical thickness and in aging wild‐type mice also led to cancellous bone gain owing to amplified bone formation rates. PTH‐induced bone gain was blunted at all doses and skeletal sites in Sost overexpressing and deficient mice owing to attenuated bone formation rates, whereas bone resorption was not different from that in PTH‐treated wild‐type controls. These data suggest that suppression of the bone formation inhibitor Sost by intermittent PTH treatment contributes to PTH bone anabolism. © 2010 American Society for Bone and Mineral Research