Osteogenic potential of cultured bone marrow stromal cells transfected with transforming growth factor β1 genein vitro

Summary To study the osteogenic potential of cultured bone marrow stromal cells (BMSCs) transfected with transforming growth factor β₁ (TGF-β₁) genein vitro, cultured BMSCs were transfected with the complexes of pcDNA₃-TGF-β₁ and Lipofectamine Reagentin vitro. The cell proliferation was detected by MTT method and the morphological features of transfected BMSCs was observed. ALP stains and PNP method were used to measure ALP activity. In addition, the collagen type I propeptides and mineralized matrixes were examined by immunohistochemical staining and tetracycline fluorescence labeling respectively. The morphological and biological characters of the transfected BMSCs were similar to those of osteoblasts and the cell proliferation was promoted. The cell layer displayed strong positive reaction for ALP stains and immunohistochemical staining. ALP activity and collagen type I expression increased remarkably after transfection. Mineralized matrixes formed earlier and more in transfected BMSCs as compared with control group. It is concluded that transfecting with TGF-β₁ gene could promote the osteogenic potential of cultured BMSCs.     

胃肠道间质瘤的临床病理及免疫组化表达

目的：观察胃肠道间质肿瘤(gastrointestinal stromal tumor,GIST）临床病理特征及免疫组化表达,为临床诊断、治疗提供可靠依据。方法：采用常规病理切片HE染色,光镜观察,并行S－P法免疫组化Vimentin、CD34、SMA、Desmin、NF、S-100、EMA等标记物标记26例GIST。结果：26例GIST中,良性7例,恶性10例,低度恶性9例;发生于胃14例,小肠5例,回盲部2例,大肠和直肠5例;免疫组化Vimentin、CD34在瘤细胞中呈强阳性表达（阳性表达率分别为96．2％和84．6％）,肌源性和神经源性标记物SMA、S－100呈散在细胞阳性表达（阳性表达率分别为26．9％和15．4％）,而NF、EMA、Desmin均呈阴性表达。结论：GIST是一种源于原始间充质细胞的非定向分化的肿瘤,可伴有神经源性或／和肌源性分化,组织学上常以梭形细胞或上皮样细胞为主,具有类似肌源性或神经源性肿瘤的形态;临床手术切除前不易诊断,病理组织形态上易误诊为其它类型梭形细胞肿瘤;CD34和Vimentin对本瘤病理诊断具有较高的价值。     

脂肪因子Chemerin对骨代谢影响的研究进展

现代社会肥胖人群逐年增多,肥胖相关代谢性骨病的患者也日益增多,严重危害人群健康。Chemerin作为一种脂肪因子,主要由脂肪细胞和肝细胞分泌,在循环中水平高于经典脂肪因子如Leptin和Adiponectin,对全身代谢和免疫发挥重要作用。近年研究发现,肥胖状态下伴随着外周脂肪组织的增多,骨髓内脂肪组织也急剧增加。骨髓内脂肪细胞分泌Chemerin,通过旁分泌或自分泌方式抑制成骨细胞分化和促进破骨细胞分化以及影响骨髓腔微环境。本文将系统回顾国内外有关Chemerin与骨代谢相关研究的文献,并探讨Chemerin对骨代谢的影响及其机制。     

人脐血间质干细胞条件培养基在受损成骨细胞中的作用与机制

目的探讨线粒体活性在人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,HUMSCs)条件培养基修复受损成骨细胞中的作用及其机制。方法提取、培养HUMSCs及制作其条件培养基,建立成骨细胞氧化损伤模型。实验分组:无血清DMEM培养基组、晚期氧化蛋白(advanced oxidation protein products,AOPPs)刺激组、常规培养基组、条件培养基组,培养1 h后用流式细胞仪以及Mito Tracker Red分析成骨细胞凋亡率、线粒体活性,并用蛋白-凝胶成像法(western-blot,WB)分析Caspase-3凋亡蛋白表达差异,从而评价HUMSCs条件培养基对遭受氧化损伤的成骨细胞的保护、修复、逆转作用,以及线粒体在其中发挥的作用及其机制。结果HUMSCs条件培养基可以增加氧化应激作用下成骨细胞内线粒体活性;HUMSCs条件培养基可降低成骨细胞凋亡率,减少Caspase-3凋亡蛋白表达。结论HUMSCs条件培养基通过缓解氧化应激(oxidative stress,OS)导致的线粒体功能与结构损伤,并下调Caspase-3凋亡蛋白的表达,抑制OB凋亡。     

miR-21对骨质疏松小鼠骨髓基质细胞增殖的影响

目的探讨miR-21对骨质疏松小鼠骨髓基质细胞(BMSCs)增殖的影响。方法采用双侧卵巢切除法构建骨质疏松小鼠模型(VOX),分离、培养、纯化小鼠BMSCs并采用si PORT Neo FX转染pre-miR-21、pre-miR-negative control(pre-miR-NC)、antmiR-21、ant-miR-negative control(ant-miR-NC)并进行RT-PCR验证,MTT法检测小鼠BMSCs增殖情况、茜素红与碱性磷酸酶染色法检测小鼠BMSCs成骨能力、Western-blotting检测细胞增殖、成骨分化相关蛋白水平。结果骨质疏松症小鼠BMSCs中miR-21相对表达水平低于Ctrl组(P0.05),OVX-pre-miR-21组BMSCs中miR-21相对表达水平、细胞增殖、PCNA水平、Ki-67水平、ALP染色程度、ALP活性、茜素红染色程度、Runx2水平、Osterix水平均高于OVX-pre-miR-NC组(P0.05),OVX-premiR-NC组BMSCs中miR-21相对表达水平、细胞增殖、PCNA水平、Ki-67水平、ALP染色程度、ALP活性、茜素红染色程度、Runx2水平、Osterix水平均显著低于Ctrl-pre-miR-NC(P0.05); OVX-ant-miR-21组BMSCs中miR-21相对表达水平、细胞增殖、PCNA水平、Ki-67水平、ALP染色程度、ALP活性、茜素红染色程度、Runx2水平、Osterix水平均显著低于OVX-ant-miR-NC组(P0.05),OVX-ant-miR-NC组BMSCs中miR-21相对表达水平、细胞增殖、PCNA水平、Ki-67水平、ALP染色程度、ALP活性、茜素红染色程度、Runx2水平、Osterix水平均显著低于Ctrl-ant-miR-NC(P0.05)。结论提高miR-21表达水平可促进骨质疏松小鼠BMSCs增殖能力与成骨分化能力。     

卵巢硬化性间质瘤1例

患者女,27岁,已婚,孕0产0,因“体检发现右侧卵巢肿物18天”入院;既往月经紊乱4年余。妇科查体:阴道少量白色分泌物,宫颈中度糜烂。实验室检查:雌二醇75.69 pg/ml,孕酮0.95 ng/ml。MRI:右侧附件区见2.1 cm×3.2 cm×3.5 cm团块状异常信号,界清,T1WI呈等信号,脂肪抑制T2WI呈等、稍高、高混杂信号(图1A),周边以等信号为主,呈乳头状、梳状深入中央高信号区,高信号区内见团块状、结节状等信号灶,形成“湖岛”征;DWI(b=1000 s/mm 2)呈稍高信号;增强扫描动脉期病灶明显不均匀强化(图1B),静脉期呈渐进性明显强化(图1C),盆腔少量积液。     

Giant hepatic extra-gastrointestinal stromal tumor treated with cytoreductive surgery and adjuvant systemic therapy: A case report and review of literature

BACKGROUNDPrimary extra-gastrointestinal stromal tumors (E-GIST) of the liver are rare. The clinical presentation may range from asymptomatic to bleeding or manifestations of mass effect. Oncologic surgery followed by adjuvant therapy with imatinib is the standard of care. However, under specific circumstances, a cytoreductive approach may represent a therapeutic option. We describe herein the case of an 84-year-old woman who presented with a tender, protruding epigastric mass. Abdominal computed tomography scan revealed a large, heterogeneous mass located across segments III, IV, V, and VIII of the liver. The initial approach was transarterial embolization of the tumor, which elicited no appreciable response. Considering the large size and central location of the tumor and the advanced age of the patient, non-anatomic complete resection was indicated. Due to substantial intraoperative bleeding and hemodynamic instability, only a near-complete resection could be achieved. Histopathology and immunohistochemical staining confirmed the diagnosis of primary E-GIST of the liver. Considering the risk/benefit ratio for therapeutic options, debulking surgery may represent a strategy to control pain and prolong survival.CASE SUMMARYHere, we present a case report of a patient diagnosed with E-GIST primary of the liver, which was indicated a cytoreductive surgery and adjuvant therapy with imatinib.CONCLUSIONE-GIST primary of the liver is a rare conditional, the treatment is with systemic therapy and total resection surgery. However, a cytoreductive surgery will be necessary when a complete resection is no possible.     

小鼠胸腺基质细胞株MTECB的细胞学特性

用免疫组织化学和透射电镜技术对本室所建立的小鼠胸腺基质细胞株ＭＴＥＣＢ进行了细胞学鉴定。免疫组织化学观察显示：４０ｋＤ、４５ｋＤ角蛋白和波形蛋白呈现强阳性，５２ｋＤ，５４ｋＤ，５８ｋＤ角蛋白和Ｓ－１００蛋白呈现阴性。电镜下见到，具有桥粒和张力丝结构，含较多的囊泡和髓样体，发达的高尔基复合体和线粒体。另外ＭＴＥＣＢ还表现为酸性磷酸酶、碱性磷酸酶、非特异性酯酶弱阳性。据此我们认为ＭＴＥＣＢ为上皮细胞来     

骨髓基质细胞和内皮细胞联合种植构建组织工程化骨的初步研究

为阐明骨髓基质细胞和内皮细胞联合种植在生物复合材料上构建组织工程化骨 ,在促进成骨过程、加速局部血管生成中的作用 ,将大鼠骨髓基质细胞和人脐静脉内皮细胞联合种植在左旋聚乳酸 /β-磷酸三钙 (PL L A/β- TCP)多孔复合支架材料上作为实验组 ,仅种植骨髓基质细胞在该材料上作为对照组。将此种有活细胞的复合材料种植于裸鼠大腿后侧肌袋内 ,分别于种植后 1、4、8、12、16周处死动物 ,取出复合材料 ,行组织学检测 ,并用图像处理技术计算实验组和对照组的成骨面积百分比和材料面积百分比 ,观察其变化规律。结果显示 :实验组成骨面积百分比增加比对照组明显加快 ,而材料面积百分比减少比对照组迅速 ;其内毛细血管网形成情况也要明显优于对照组。表明骨髓基质细胞和内皮细胞联合种植于支架材料构建组织工程化骨不仅有利于成骨作用 ,促进新生骨区内部血管网形成 ,还有利于复合材料的降解吸收 ,这在构造组织工程化骨过程中具有重要意义。