新型活性修饰聚乳酸组织工程骨的体外成骨研究

[目的]探讨氨等离子体改性、酰胺键接枝甘氨酸-精氰酸-甘氨酸-天冬氨酸-丝氨酸(GRGDS)短肽的活性修饰方法对消旋聚乳酸(PDLLA)组织工程骨体外成骨能力的影响.[方法]制备圆片状直径8 mm、厚1 mm的PDLLA三维多孔支架,分为三组:表面氨基化PDLLA(aminated PDLLA,A/PDLLA,A组),接枝肽A/PDLLA(peptides conjugated A/PDLLA,PA/PDLLA,PA组),以未经处理的PDLLA(P组)作为对照组.实时荧光定量聚合酶链式反应(RT-qPCR)检测符组材料上骨髓间允质干细胞(BMSCs)、骨钙素(OCN)、Ⅰ型胶原(Col-Ⅰ)、碱性磷酸酶(ALP)、骨形态发生蛋白-2(Bmp-2)和骨桥蚩白(OPN)mRNA的表达变化并进行ALP活性测定和钙黄绿素矿化荧光染色.[结果]qPCR结果示:第3 d,PA组各基因表达倍数均较P组高;除OCN外的各基因表达也均较A组高.第7 d,A组OCN(13.13±1.28)、Col-Ⅰ(23.71±6.51)和OPN(27.4±7.17)mRNA表达倍数为最高,PA组次之.第14 d,多数基因表达上调,A组和PA组表达倍数较对照组明显增高.PA组OCN(49.21±7.03)、ALP(24.26±3.41)和BMP-2(11.82±2.38)mRNA的表达较A组高,差异有统计学意义(P＜0.05).ALP活性检测结果示:A组和PA组ALP活性持续升高,P组活性第21 d较第14 d时略有下降.第7 d,A组和PA组之间ALP活性比较无统计学意义(P＞0.05),但均高于P组(P＜0.01);第14 d和第21 d,三组间ALP活性两两比较均有统计学意义,ALP活性PA组＞A组＞P组.钙黄绿素矿化荧光染色观察示,骨支架上钙盐沉积与ALP的活性变化趋势相平行.[结论]氨等离子体改性能够促进聚乳酸组织工程骨支架上BMSCs早期向成骨细胞分化,改性后接枝GRGDS肽的新型活性修饰PDLLA具有更好的促BMSCs体外成骨能力.     

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

目的 探讨灌注培养对骨髓基质干细胞(BMSCs)在大段材料上增殖与分布的影响.方法 在大段多孔磷酸三钙材料上种植转染了增强型绿色荧光蛋白基因的大鼠BMSCs(eGFP-BMSCs),分别采用灌注式生物反应器(实验组)和静止培养法(对照组)进行培养.培养7、28 d行扫描电镜观察;培养7、14 d行荧光显微镜观察;动态临...     

同种异体与自体血清对骨髓基质干细胞的增殖效应

背景：传统的细胞培养方法大多采用胎牛血清作为营养支持,其存在传播疾病的风险和免疫排斥反应。寻找胎牛血清替代物并建立规范、安全、高效的成人骨髓基质干细胞体外培养体系刚刚起步,有待进一步研究。 目的：探讨AB血清、自体血清替代胎牛血清体外培养人骨髓基质干细胞的效果。 方法：从成人骨髓穿刺液中分离人骨髓基质干细胞,分别用含体积分数10%AB血清、自体血清和胎牛血清的基础培养基培养人骨髓基质干细胞,取第3代细胞进行相关指标检测。 结果与结论：相差显微镜下见AB血清组细胞最先达到汇合;流式细胞仪检测细胞表面抗原,各组均符合骨髓干细胞表型,纯化程度良好;Alamar Blue法显示AB血清组平均荧光强度最高,促增殖效率最高;Annexin V-FITC法显示各组细胞凋亡率均在5%以下,生长状态良好;碱性磷酸酶染色、钙结节和油红O染色结果显示,各组细胞均保持了成骨、成脂分化能力。AB血清对人骨髓基质干细胞的增殖作用较胎牛血清、自体血清明显增强。AB血清有望替代胎牛血清建立符合骨组织工程临床应用要求的体外成人骨髓基质干细胞培养体系。 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

CLIA在肿瘤标志物检测中的应用

化学发光技术是指当基态分子吸收化学反应中释放的能量跃迁到激发态,处于激发态的分子以光辐射的形式返回基态时产生的光称为化学发光。基于化学发光强度和被测物含量之间的关系建立的分析方法叫化学发光分析法,它具有灵敏度高,线性范围宽,仪器简单、操作简便等特点,已广泛地应用于环境、临床、食品和工业分析中。1977年,Tsuji等[1]基于放射免疫分析的基本原理,将酶的化学发光与免疫反应结合起来,发展了化学发光免疫分析(Chemiluminescence Immunoassay,CLIA)方法。它是将发光物质或酶标记在抗原或抗体上,免疫反应结束后,加入氧化剂或酶底物而发光,通过测量发光强度,根据标准曲线测定待测物的浓度。CLIA的主要优点是灵敏度高、线性范围宽、标记物的有效期长、无放射性危害、可实现全自动化等。CLIA具有强大的生命力,在国际和国内倍受临床检验和生物医学工作者的关注。1化学发光免疫分析基本原理化学发光免疫分析包含两个部分,即免疫反应系统和化学发光分析系统[2]。化学发光(Chemiluminescence,CL)是化学物质在特定化学反应中产生的光辐射。通过高能中间体的分解在化学反应中激发单态分子形成,分子被...     

Morel-Lavallée损伤的临床病理特点及治疗现状

Morel-Lavallée损伤是一类严重的闭合性软组织脱套伤,常见于高能量剪切暴力损伤,可发生在身体不同部位,经常伴随骨盆、髋臼等各类骨折的发生。尽管该损伤发生率不高,但这类损伤十分特殊,早期隐蔽性较强。临床外科医师对此类损伤缺乏足够的重视,往往处理不及时或处理不当,导致皮肤软组织坏死、感染等诸多并发症。目前,Morel-Lavallée损伤没有标准的分类系统,主要基于MRI影像学表现,但对于指导临床治疗及预后无实际意义。其治疗方案多样,如局部加压包扎、经皮穿刺抽吸、有限切开清创冲洗、负压吸引、置管引流等,效果不一。就Morel-Lavallée损伤的研究现状予以综述。     

自体髌腱与四股腘绳肌腱移植重建前交叉韧带安全性的Meta分析

目的 评价自体髌腱与4股腘绳肌腱移植关节镜下单束重建前交叉韧带(ACL)的安全性.方法 计算机检索CENTRAL、PubMed、EMBASE、中国生物医学文献数据库等,获取自体髌腱与4股腘绳肌腱移植重建ACL的随机对照试验(RCT),根据Cochrane图书馆推荐的RCT偏倚评估标准对纳入研究的方法学质量进行评估,并使用RevMan 5.0统计学软件对结果进行Meta分析,比较自体髌腱与4股腘绳肌腱移植重建ACL的移植失败率、伸膝缺失度、膝前痛等的差异.结果共纳入14篇文献,包括1232例患者.自体髌腱组与自体4股腘绳肌腱组的移植失败率在平均随访2～3年(RR=0.69,95%CI 0.34～1.38,P=0.29)、5年(RR=0.37,95%CI0.09～1.55,P=0.18)、10年(P=0.94)及合并后(RR=0.71,95%CI 0.39～1.31,P=0.28)差异均无统计学意义.两组伸膝缺失度≥5°的例数比较差异无统计学意义(RR=1.63,95%CI0.85～3.16,P=0.14).两组膝前痛随访2年差异有统计学意义(RR=2.04,95%CI 1.34～3.09,P=0.0008),而随访5年(RR=3.00,95%CI 0.92～9.79,P=0.07)及随访10年(P=0.737)差异均无统计学意义.两组膝跪痛或不适随访2年差异有统计学意义(RR=1.97,95%CI 1.01～3.85,P=0.05),随访5年(RR=1.51,95%CI 0.95～2.39,P=0.08)及随访10年(P=0.342)差异无统计学意义.结论自体髌腱与4股腘绳肌腱移植失败率及膝关节被动伸直缺失的发生率相似.自体髌腱移植随访早期(平均随访2～3年)膝前痛、膝跪痛或不适的发生率较高,但随着时间的延长(5～10年),两者趋于一致.

Abstract：

Objective To compare clinical outcomes of patellar tendon and 4-strand hamstring autografts in single-bundle endoscopic reconstruction of anterior cruciate ligament(ACL). Methods The digital databases(PubMed,Embase,Cochrane Central Register of Controlled Trials,CBM)were searched for randomized controlled trials (RCTs)comparing patellar tendon with hamstring autografts for reconstruction of ACL.In addition.the reference lists from related original studies and review articles were hand-searched.The Cochrane Collaboration's tool for assessing risk of bias and Cochrane collaboration's RevMan 5.0 software were used for assessing trial methodological quality and data analyses. Results Fourteen RCTs were included.There were 1232 participants who met inclusion criteria in all studies.Meta-analyses showed that no statisticallv significant differences were found in graft failure rate after single-bundle reconstruction of the ACL at mean follow-ups of 2 to 3 years (RR=0.69,95%C/0.34 to 1.38,P=0.29,of 5 years (RR=0.37,95%CI 0.09 to 1.55,P=0.18),of 10 years(P=0.94)or when 3 subgroups combined(RR=0.71,95%CI 0.39 t0 1.31.P=0.28).The diffe-rence in≥5°extensor deficit was not statistically significant between the 2 groups(RR=1.63.95%CI 0.85 to 3.16,P=0.14).Anterior knee pain and kneeling pain or discomfort showed significantlv differences between the 2 groups at a mean follow-up of 2 years(P<0.05),but did not at follow-ups of 5 years and 10 years(P>0.05). Conclusions In single-bundle endoscopic ACL reconstruction.patellar tendon and 4-strand hamstring autografts have similar rates of graft failure and knee motion deficits.The patellar tendon autograft may have higher rates of anterior knee pain and kneeling pain or discomfort at an early stage after operation than the 4-strand hamstring autografi,but there may be no such differences on a long-term basis.     

关节镜下自体4股腘绳肌腱双束与单束重建前交叉韧带后膝关节稳定性及功能评分的meta分析

[目的]评价关节镜下自体4股腘肌腱双束与单束重建前交叉韧带的临床效果.[方法]计算机检索CENTRAL、PubMed、Embase、CBM等数据库,获取自体4股腘绳肌腱移植双束与单束重建前交叉韧带的随机对照试验,根据Cochrane协作网推荐的随机对照试验偏倚评估工具对纳入研究的方法学进行评估,并使用RevMan5.0对数据进行Meta分析.[结果]共纳入10篇文献,693例前交叉韧带损伤患者纳入Meta分析,结果显示:双束重建组术后KT2000或KT1000测量值与单束重建组的差异有统计学意义(SMD=-0.27,95% CI(-0.53,-0.01),P=0.04);轴移试验阴性(RR=1.35,95% CI(1.16,1.56),P<0.000 1)、弱阳性(RR=0.37,95% CI(0.18,0.74),P=0.005)、阳性(RR =0.17,95% CI (0.06,0.66),P=0.01)的差异均有统计学意义.功能评分中IKDC主观评分(SMD=0.03,95% CI(-0.21,0.28),P=0.79)和Lysholm评分(SMD=-0.25,95% CI(-0.53,0.02),P=0.07)均无统计学意义.[结论]自体4股腘绳肌腱双束重建前交叉韧带在恢复膝关节的前直向稳定性和旋转稳定性方面优于单束重建;但功能评分相似.     

HX-21自动细菌鉴定/药敏分析系统简介

概述　　合肥恒星科技开发有限公司是我国安徽省规模最大的一家专业从事医疗检验产品研发、生产的高新技术企业。公司创建于 1993年 ,原属省科委 ,目前有员工 6 0余名 ,大专以上学历占百分之九十以上。公司以高校为依托、以科研机构为技术联盟 ,实现了资源共享、共同开发和长期的合作关系 ,单位有 :中国科技大学生物技术研究所、浙江大学生命科学院、安徽医科大学附属医院、安徽省立医院 (临床实验基地 ) ;一批青年博士、硕士组成的研发队伍是公司长期稳定发展的人才后盾和可靠保障。HX— 2 1自动化细菌鉴定 /药敏分析系统为安徽省“十五”…     

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.     

自体髌腱与四股腘绳肌腱移植重建前交叉韧带安全性的Meta分析

Objective To compare clinical outcomes of patellar tendon and 4-strand hamstring autografts in single-bundle endoscopic reconstruction of anterior cruciate ligament(ACL). Methods The digital databases(PubMed,Embase,Cochrane Central Register of Controlled Trials,CBM)were searched for randomized controlled trials (RCTs)comparing patellar tendon with hamstring autografts for reconstruction of ACL.In addition.the reference lists from related original studies and review articles were hand-searched.The Cochrane Collaboration's tool for assessing risk of bias and Cochrane collaboration's RevMan 5.0 software were used for assessing trial methodological quality and data analyses. Results Fourteen RCTs were included.There were 1232 participants who met inclusion criteria in all studies.Meta-analyses showed that no statisticallv significant differences were found in graft failure rate after single-bundle reconstruction of the ACL at mean follow-ups of 2 to 3 years (RR=0.69,95%C/0.34 to 1.38,P=0.29,of 5 years (RR=0.37,95%CI 0.09 to 1.55,P=0.18),of 10 years(P=0.94)or when 3 subgroups combined(RR=0.71,95%CI 0.39 t0 1.31.P=0.28).The diffe-rence in≥5°extensor deficit was not statistically significant between the 2 groups(RR=1.63.95%CI 0.85 to 3.16,P=0.14).Anterior knee pain and kneeling pain or discomfort showed significantlv differences between the 2 groups at a mean follow-up of 2 years(P<0.05),but did not at follow-ups of 5 years and 10 years(P>0.05). Conclusions In single-bundle endoscopic ACL reconstruction.patellar tendon and 4-strand hamstring autografts have similar rates of graft failure and knee motion deficits.The patellar tendon autograft may have higher rates of anterior knee pain and kneeling pain or discomfort at an early stage after operation than the 4-strand hamstring autografi,but there may be no such differences on a long-term basis.