不同浓度5-溴脱氧尿嘧啶核苷标记山羊骨髓基质干细胞的体外研究

Objective To explore the feasibility of labeling and tracing in vitro goat bone marrow mesenchymal stem cells (BMSCs) by bromodeoxyuridine (BrdU) on the basis of investigation of its optimal concentration, incubating time and cytotoxicity. Methods A healthy goat, aged 10 months old, male, weighing 32 kg, was used in this study. Bone marrow was aspirated. BMSCs were isolated and cultured using the adherence method in vitro. The fourth passage of BMSCs (P4) were incubated with BrdU at 5, 10, 15, 20 μmol/L as 5, 10, 15, 20 μmol/L BMSC groups. Cells were not labeled by BrdU as negative control. The following parameters were measured: induction, differentiation and determination of goat BMSCs; the optimal mass concentration and incubation time of 5-BrdU labeling; cell positive rate at 12, 24, 48 and 72 hours in each group using immunofluoreseenee; the cell survival rate after various concentrations of BrdU ladling by trypan-blue exclusion. Results The morphology of the primary and passage goat BMSCs was fusiform in shape. Goat BMSCs could differentiate into osteoblasts and chondrocytes following induction. BMSC nucleus showed green fluorescence under fluorescence microscope after being labeled by BrdU. The mean labeling rate increased with the increase in the concentration and incubation time of BrdU, and reached to (93.32± 3.25)% after incubation in 15 μmol/L, BrdU for 48 hours. There were no significant differences between 15 μmol/L BrdU for 72 hours, 20 μmol/L BrdU for 48 hours and 72 hours (P > 0.05), or between the other groups or time points (P < 0.05). The labeling rate of the blank control group was 0. The cell survival rate was all above 90% (P > 0.05). Conclusions BrdU can be used as a labeling marker for goat BMSCs. When the concentration is 15 μmol/L and the incubation time is 48 hours, the optimal labeling effect can be achieved. Goat BMSCs labeled with BrdU is of high efficiency and safety.     

双极电凝器在普外科手术中的应用

目的　探讨双极电凝器在普外科手术中的应用价值。方法　在 5 6 2例普外科手术中应用国产“新迪”牌XDS 1型双极电凝器进行止血操作 ,依病变部位等具体情况选择相应的操作方法。结果　全组病例应用双极电凝止血均获成功 ,无一例发生因应用双级电凝而造成的术后大出血。结论　普外手术中应用双极电凝止血具有操作简便、安全可靠、并发症少的优点 ,特别是在深部组织和精细组织的止血中有较高的应用价值 ,值得推广     

基于Unigraphics NX和Mimics软件的股骨远端骨折三维虚拟手术研究

目的 利用Unigraphics NX和Mimics软件进行股骨远端骨折虚拟手术,为术前精确设计手术方案提供真实可靠的依据.方法 选取6例股骨远端骨折患者,利用Mimics 10.01软件对股骨远端骨折患者的CT扫描数据进行三维重建,Unigraphics NX软件对手术器械进行实体建模.利用U-nigraphics NX软件系统仿真模拟股骨远端骨折微创内固定系统钢板固定手术和逆行髓内钉固定手术,进行股骨远端骨折患者术前的虚拟手术研究,初步指导临床应用.结果 仿真手术过程真实逼真,立体可视化效果好,并可预测骨折复位状念及手术效果,微创内固定系统钢板对位合适,髓内钉对位对线准确,螺钉进针点安全可靠.结论 利用Unigraphics NX和Mimics软件进行虚拟手术具有较强的可视效果,对提高股骨远端骨折手术的安全性和准确性具有重要作用.     

正常人足踝部有限元模型的构建研究

目的 研究足踝部有限元模型的构建方法,为足踝部生物力学提供一个数字化研究平台. 方法 获取一名30岁男性志愿者的右足螺旋CT扫描图像,利用Mimics软件重建出足踝部28块骨骼及外围软组织的三维结构,通过Solidworks处理后导入Ansys的Workebench模块,建立足踝部三维有限元模型.在模型中还通过解剖和文献数据模拟建立各关节软骨、韧带、跖筋膜、小腿骨间膜,然后模拟踝关节正常站立状态下受力,选择相应边界条件进行加载分析. 结果建立了一个包含骨与外围软组织等复合结构的正常人体右足三维有限元模型,相埘客观地反映了人体足踝部的基本解剖结构和力学特性.模拟人体站立状态垂直加载600 N载荷于胫骨下端的上截面,踝关节胫骨下关节面应力主要分布于中部及前外侧,最大应力为3.97 MPa,平均接触应力为1.52 MPa,接触面积为343.6 mm~2. 结论本研究所建立的止常人体足躁部三维有限元模型经验证结果可靠,可进一步用于足踝部损伤的研究.     

胫骨近端截骨面与胫骨假体的数字化形态学研究

目的 利用数字化技术获取健康成人正常胫骨近端截而及假体的参数参考值,探讨适合不同人种且具有性别差异的胫骨假体设计.方法 对55名志愿者中的92侧正常膝关节(男42侧,女50侧)进行螺旋CT扫描,将断层扣描数据导入Mimics10.1软件建立膝关节三维数字模型,然后将模型导入Geomagic Studio 8软件系统对胫骨近端模型进行模拟截骨、形态学测量,分析不同性别胫骨近端截面的形态学差异.再将获得的模型与通过激光扫描获得的假体(PFC和Nexgen)三维模型进行模拟装配、参数测量.结果 胫骨近端截骨面的前后径(AP)为(49.87 ±3.90)mm、内外侧横径(ML)为(73.50±5.60)mm,AP与ML呈正相关.男性胫骨平台AP为(53.24±4.10)mm、ML为(75.85±3.50)mm,均比女性[AP为(47.61±3,70)mm,ML为(67.68±2.60)mm]大.内侧前后径(MAP)比外侧前后径(LAP)大,男性平均大(3.90±2.90)mm,女性平均大(3.70±2.70)mm.PFC假体(0.75±0.05)和Nexgen假体(0.69±0.03)的AP/MP比值与本研究正常成人(0.68±0.03)相比,差异均有统计学意义(P<0.05).结论 不同性别及人种的胫骨截面形态存在显著差异,为提高人工膝关节胫骨假体的覆盖率,胫骨假体设计及临床操作均要考虑不同人种和性别的形态学特点.     

灌注培养对骨髓基质干细胞在大段材料上增殖与分布的影响

Objective To study the role of perfusion bioreactor in proliferation and distribution of rat bone marrow stromal cells (BMSCs) in a large-scale scaffold. Methods SD rat BMSCs transfected with enhanced green fluorescent protein (eGFP) (eGFP-BMSCs) were planted in large-scale porous β-tricalcium phosphate (β-TCP) scaffolds. In the dynamic perfusion culture group, the scaffold with eGFP-BMSCs was continuously cultured in our self-designed three-dimensional perfusion bioreactor for 7, 14 and 28 days. In the static culture group, the scaffold was put into a medium reservoir without perfusion for 7, 14 and 28 days.Proliferation and distribution of the cells in the scaffold were examined by scanning electronic microscopy (SEM), fluorescence microscopy (FM), measuring daily glucose consumption, and counting eGFP-BMSCs in each layer. Results SEM and FM showed that eGFP-BMSCs distributed and proliferated throughout the scaffold in dynamic perfusion culture, but distributed and proliferated only in the peripheral pores of the scaffold in static culture. The daily glucose consumption in both groups increased with time. Cell proliferation reached the plateau phase after culture for 14 days in the static culture group, but after culture for 21 days in the perfusion culture group. The rate and margin of increase were much more evident in the perfusion culture group. On day 28, glucose consumption in the perfusion culture group was 36. 33 ± 3. 14 mg/d, 3. 7 times as large as that in the static culture group (9. 82 ± 1. 33 mg/d). The eGFP-BMSCs counting revealed there were no significant differences in cell number between layers of the scaffold on days 7 and 28 d ( P > 0. 05), but there were significant differences on day 14 in the perfusion culture group ( P < 0. 05); while in the static culture group, there were significant differences in cell number between layers of the scaffold ( P < 0. 05),with most of the cells assembled at the bottom of the scaffold.Conclusion Our self-designed three-dimensional perfusion bioreactor may help eGFP-BMSCs proliferate and distribute uniformly in a large-scale porous scaffold.     

中药贴敷疗法治疗小儿肺炎喘嗽的临床观察

肺炎喘嗽是儿科常见病,临床以发热、咳嗽、痰壅、气急、鼻煽为主要症状。由于小儿存在服药困难的特点,近年来,作为中医外治法的中药贴敷疗法在小儿肺炎喘嗽的治疗中显示出了一定优势,疗效肯定。     

左炔诺孕酮宫内缓释系统对子宫内膜增生过长的疗效分析

目的 评估左炔诺孕酮宫内缓释系统(LNG-IUS)对子宫内膜增生过长患者的治疗效果和可接受性.方法 观察25例子宫内膜增生过长患者LNG-IUS放置前后的症状、组织学改变、不良反应等.结果 所有患者放置LNG-IUS后月经量均明显减少,贫血逐渐改善,LNG-IUS放置后6个月子宫内膜明显变薄,表现为子宫内膜腺体数目明显减少,腺腔变小,腺体发育不良,间质细胞有明显的蜕膜样变.25例患者均对疗效表示满意,不良反应主要是不规则少量阴道出血或点滴状出血,无患者中途取出LNG-IUS.结论 LNG-IUS对于治疗子宫内膜增生过长的患者是有效的,良好咨询有助于提高患者的可接受性.     

MR3.0T磁敏感技术在隐匿性脑外伤中的临床应用价值

目的 探讨3.0T磁敏感加权成像(SWI)技术在隐匿性脑外伤中的临床应用价值.方法 搜集脑外伤患者24例,均行常规T1WI、T2WI、FLAIR及SWI.结果 24例头部CT平扫、MR平扫T1WI、T2WI、FLAIR像阴性,SWI清楚地显示了脑内微出血灶.结论 在隐匿性脑外伤患者中,SWI较常规扫描序列更多的显示微出血状况,为临床提供更多的诊断信息,可作为隐匿性脑外伤常规扫描序列.     

miR-21真核表达载体的构建及其对胃癌细胞增殖与凋亡的影响

目的探讨人微小RNA-21（miR-21）重组表达质粒对人胃癌SGC-7901细胞增殖与凋亡的影响及可能机制。方法设计合成靶向miR-21的互补双链DNA,退火后与真核表达载体pPG—miR．EGFP克隆连接,构建pPG—miR-21-EGFP重组表达质粒,利用脂质体LipofectamineTM2000将鉴定正确的重组表达质粒pPG—miR-21．EGFP转染人胃癌SGC-7901细胞,荧光显微镜下观察转染效率。实验分为3组：空白对照组、空载体组、重组表达质粒组。以RT—PCR检测转染前后胃癌细胞miR-21的表达水平。MTY法评价重组表达质粒抑制肿瘤细胞生长的效果,流式细胞术及Hoechst33258法检测转染后胃癌SGC-7901细胞周期的分布和凋亡。Westernblot法、免疫细胞化学法分别检测转染前后PTEN、PCNA、PDCD4、bcl-2、cyclinD1蛋白表达的水平。结果酶切及测序鉴定证实成功构建重组质粒pPG—miR-21-EGFP,转染人胃癌SGC-7901细胞后,荧光显微镜下观察转染效率达75％。RT—PCR检测显示重组表达质粒组胃癌细胞miR-21的表达下调,与空白对照组和空载体组比较,差异具有统计学意义（P〈O．05）。Westernblot法、免疫细胞化学法检测结果显示重组表达质粒组胃癌细胞cyclinD1、PCNA、bcl-2蛋白的表达下调（P〈0．05）,而PDCD4、PTEN蛋白的表达上调（尸〈0．05）。重组表达质粒组的细胞生长缓慢,凋亡指数增加。流式细胞术检测结果显示重组表达质粒组G。期细胞的百分比较空白对照组增加[（69．71±0．314）％（50．1~0．331）％],S期细胞较空白对照组明显减少[（14．68~0．448）％口s（28．47~0．316）％]（P〈0．05）。结论重组表达质粒pPG．miR．21-EGFP可显著下调miR-21在胃癌SGC-7901细胞中的表达,并在-定程度上抑制胃癌细胞的增殖,促进其凋亡,使较多的细胞停留在G。期,s期细胞减少,其作用机制可能是通过下调cyclinD1、PCNA、bcl．2蛋白的表达,上调PDCD4、PTEN蛋白的表达而实现的。