自体富血小板凝胶的制备及其生长因子分析

目的 探讨不同离心方法 制备自体富血小板凝胶(autologous platelet-rich gel,APG)的方法 ,通过改变离心速度比较不同的离心力对PLT富集的影响分析全血和APG中5种生长因子浓度.方法 对13例糖尿病难治性皮肤溃疡患者行APG治疗.取11例自身外周静脉血,分别用3种离心速度制备富血小板血浆(platelet-rich plasma,PRP).A组(n=6)先以529×g离心4 min,再以854×g离心6 minB组(n=5)先以313×g离心4 min,再以1252×g离心6 min;C组(n=5)先以176×g离心5 min,再以1 252×g离心5 min.将离心后制得的PRP与凝血酶.钙剂以101混合凝固后制备APG,用于患者皮肤溃疡的治疗.采用全自动血细胞分析仪计数各组全血和PRP中PLT数量.采用酶联免疫吸附法测定全血和APG中PDGF.BB、VEGF、IGF.1、EGF和TGF.a1 5种生长因子浓度.结果 A组PRP中PLT数量为(779.67±352.39)×109/L,较全血的(263.50±76.63)×109/L提高(2.98±1.42)倍,差异有统计学意义(P＜0.05);PLT回收率为51.5%±22.2%.B组PRP中PLT数量最高,为(1363.80±919.74)X 109/L,较全血的(232.80±127.99)×109/L提高(5.91±2.04)倍,差异有统计学意义(P＜0.05);PLT回收率为75.2%±21.0%,明显高于A组(P＜0.05).全血和APG中PDGF-BB、EGF、IGF-1以及TGF-a.浓度分别为(145.94±133.24)、(503.81±197.86)pg/mL,(160.73±71.10)、(265.95±138.43)pg/mL,(14.54±35.34)、(110.56±84.36)ng/mL,(3.31±2.27)、(5.67±4.80)ng/mL,差异有统计学意义(P＜0.05);VEGF浓度升高,两者间差异无统计学意义(P＞0.05).对数转换后的PLT数量与PDGF-BB、TGF-a1浓度成正相关,相关系数r分别为0.627和0.437(P＜0.05).13例患者共行18次APG治疗,其中9例治疗12周溃疡愈合,愈合率为69.2%;10例窦道愈合,愈合率为83.3%.结论 以313×g离心4 min,再以1252×g离心6 min是制备PRP的最佳方法 ;APG中生长因子浓度高于全血;PLT数量与PDGF-BB、TGF-a1浓度成正相关.     

血管瘤中血管内皮生长因子表达的定量研究

目的观察血管内皮生长因子(VEGF)在血管瘤中的表达,并探讨VEGF在血管瘤中的表达水平与DNA倍体、细胞增殖活性及血管瘤侵袭的关系.方法应用流式细胞仪及免疫荧光技术检测几种类型的血管瘤组织中VEGF表达量荧光指数FI、DNA含量(DNA指数,DI)及细胞增殖活性(增殖指数,PI).结果在血管瘤中多数为DNA异倍体、各类血管瘤FI、DI、PI值之间差异无显著性意义(P＞0.05);包膜完整组与无包膜或包膜不完整组之间FI值差异有显著性意义(P＜0.01);异倍体与二倍体之间FI值差异有显著性意义(P＜0.01);VEGF表达阳性组与VEGF表达阴性组之间PI值差异有显著性意义(P＜0.01).结论VEGF表达水平与血管瘤的侵袭性、DNA倍体及细胞增殖活性密切相关.提示阻止VEGF表达能作为治疗血管瘤的方法.     

Two kinds of posterior approach for Kümmell's disease after osteoporotic thoracolumbar fracture

To compare the surgical results of two kinds of posterior approach for osteoporotic thoracolum-bar Kummell's disease.Methods: Clinical and radiographic results of 1-seg-mental pedicle screw fixation combined with vertebroplasty (Group A, n=12) or posterior shortening osteotomy (Group B, n=16) for osteoporotic thoracolumbar Kummell's disease were analyzed retrospectively. Japanese orthopedic asso-ciation (JOA) and visual analogue scale (VAS) scores were used for clinical evaluation. Neurological status was judged by Frankel grades. X-ray was used to evaluate the radio-graphic results. Complications related to operation and de-vices were also considered. Results: The follow-up period was 12-54 months (average 29 months). Pre- and post-operative VAS were 9.3 and 3.2 in Group A, 8.9 and 2.5 in Group B, respectively. The mean JOA score at the final follow-up was significantly higher than that of pre-operation (t=5.306, P<0.001). There was no sig-nificant difference between Groups A and B (t=0.618, P>0.05). The kyphosis were corrected from preoperative 33.9°(A)/37.3°(B) to postoperative 10.3°(A)/6.5°(B), and 15.3°(A)/13.7°(B) at the final follow-up. There was a significant difference between the two groups at the final follow-up. Frankel grade was improved from grade C preoperatively to postopera-tively grade D or E in 7 cases of Group A and 5 cases of Group B, from grade D to E in 5 cases of Group A and 11 cases of Group B. The mean improvement was 1.6 and 1.7 grades for Groups A and B, respectively. There were no serious complications related to internal fixation. Conclusions: The similar clinical results can be ob-tained by the two kinds of posterior surgical methods for osteoporotic Ktimmell's disease. Posterior spinal shorten-ing is a better choice for patients with serious kyphosis combined with neurological deficit than the other.     

急性坏死性胰腺炎大鼠胰腺高迁移率族蛋白-1的表达及意义

目的探讨急性坏死性胰腺炎(ANP)大鼠模型胰腺组织高迁移率族蛋白-1(HMGB1)的表达及其意义。方法72只大鼠随机分成3组,即对照组、ANP组和正丁酸钠治疗组(治疗组)。逆行性胰胆管注射5%牛磺胆酸钠建立ANP模型。ELISA法检测血清TNF-α和IL-1β水平;RT-PCR法检测胰腺组织HMGB1 mRNA的表达,并观察其病理变化。结果ANP组血清TNF-α和IL-1β水平在ANP建模后6h达高峰,12h下降。ANP组大鼠胰腺组织HMGB1 mRNA表达水平在ANP后12h明显升高,至24h仍维持在较高水平。治疗组胰腺组织HMGB1 mRNA表达水平在ANP后12,24h明显低于ANP组(P<0.05),且同期胰腺损伤比ANP组轻(P<0.05)。建模后24h血清TNF-α和IL-1β水平ANP组与治疗组间差异无显著性。结论HMGB1作为晚期炎症因子参与了ANP的全身炎症反应。HMGB1抑制剂正丁酸钠能降低ANP大鼠胰腺组织HMGB1基因表达水平,减轻ANP胰腺组织的损伤。     

踝关节镜下治疗距骨剥脱性骨软骨炎

目的探讨踝关节镜技术在距骨剥脱性骨软骨炎治疗中的应用及效果。方法对17例距骨剥脱性骨软骨炎患者施行踝关节镜手术,在关节镜下对损伤关节面清创、搔刮及钻孔。结果随访14．54个月。踝关节Kofoed评分由术前平均56．5分提高至89．4分,17例中优6例,良8例,可2例,差1例,优良率为82．4％。结论踝关节镜下手术治疗距骨剥脱性骨软骨炎,疗效确切,具有创伤小、并发症少、恢复快等优点。     

Two kinds of posterior approach for Kümmell's disease after osteoporotic thoracolumbar fracture

To compare the surgical results of two kinds of posterior approach for osteoporotic thoracolum-bar Kummell's disease.Methods: Clinical and radiographic results of 1-seg-mental pedicle screw fixation combined with vertebroplasty (Group A, n=12) or posterior shortening osteotomy (Group B, n=16) for osteoporotic thoracolumbar Kummell's disease were analyzed retrospectively. Japanese orthopedic asso-ciation (JOA) and visual analogue scale (VAS) scores were used for clinical evaluation. Neurological status was judged by Frankel grades. X-ray was used to evaluate the radio-graphic results. Complications related to operation and de-vices were also considered. Results: The follow-up period was 12-54 months (average 29 months). Pre- and post-operative VAS were 9.3 and 3.2 in Group A, 8.9 and 2.5 in Group B, respectively. The mean JOA score at the final follow-up was significantly higher than that of pre-operation (t=5.306, P<0.001). There was no sig-nificant difference between Groups A and B (t=0.618, P>0.05). The kyphosis were corrected from preoperative 33.9°(A)/37.3°(B) to postoperative 10.3°(A)/6.5°(B), and 15.3°(A)/13.7°(B) at the final follow-up. There was a significant difference between the two groups at the final follow-up. Frankel grade was improved from grade C preoperatively to postopera-tively grade D or E in 7 cases of Group A and 5 cases of Group B, from grade D to E in 5 cases of Group A and 11 cases of Group B. The mean improvement was 1.6 and 1.7 grades for Groups A and B, respectively. There were no serious complications related to internal fixation. Conclusions: The similar clinical results can be ob-tained by the two kinds of posterior surgical methods for osteoporotic Ktimmell's disease. Posterior spinal shorten-ing is a better choice for patients with serious kyphosis combined with neurological deficit than the other.     

带蒂阴囊中隔皮瓣加盖睾丸鞘膜尿道成形术在失败尿道下裂手术中的应用

目的:评价带蒂阴囊中隔皮瓣加盖睾丸鞘膜尿道成形术在失败尿道下裂手术中的应用,了解其适应证、手术方法、术后并发症及远期效果。方法:回顾性分析2008年1月~2013年12月采用带蒂阴囊中隔皮瓣加盖睾丸鞘膜尿道成形术治疗的43例手术失败的尿道下裂病例及随访资料,描述手术适应证、手术方法,总结术后并发症及处理方法,评价远期效果。结果:平均手术时间110min。术后并发症发生率为14%(6/43),其中尿道皮肤瘘3例,尿道憩室1例,尿道口狭窄2例。术后1年随访,阴茎外观满意率为90.7%(39/43),通过随诊观察排尿评估尿道功能正常率为95.35%(41/43)。5例尿道镜检查无明显毛石形成。结论:带蒂阴囊中隔皮瓣加盖睾丸鞘膜尿道成形术是在失败尿道下裂修复术中有效的手术方式,手术方法相对简单,容易掌握,手术并发症较少,远期效果确切。     

LIN28A和LAMP1在膀胱癌细胞系中的表达及意义

目的:探讨LIN28A和LAMP1在膀胱癌细胞系中表达情况,以及两者之间的关系,推测其可能临床意义及对肿瘤进展的影响。方法:采用RT-PCR检测膀胱癌细胞系LIN28A、LIN28B和LAMP1表达,免疫荧光检测LIN28A和LAMP1二者蛋白表达定位;LIN28A敲减后通过qRT-PCR检测LAMP1的mRNA表达变化。结果:5个癌细胞系T24、UM-UC3、J82、5637和SW780和正常移行上皮细胞系SV-HUC-1均表达LIN28A,其中J82也表达LIN28B;5个癌细胞系均表达LAMP1,SV-HUC-1不表达LAMP1;LIN28A和LAMP1蛋白均定位在胞浆;LIN28A敲减后对LAMP1的mRNA表达变化无明显影响,相应蛋白变化需要进一步验证。结论:4个膀胱癌细胞系T24、5637、UM-UC3和SW780可以用于LIN28A与肿瘤相关的机制研究,而J82可用于LIN28B的机制研究。LIN28A对肿瘤细胞和干细胞的调控方面可能具有相似性,敲减后对其靶点mRNA表达量无明显影响,LAMP1蛋白可能对肿瘤细胞侵袭转移具有抑制作用。     

透明质酸受体精子的质量特性研究

目的:分析具有透明质酸受体精子的质量特征,寻找精子质量评估新指标。方法:用透明质酸包被载玻片检测活动精子-透明质酸结合率,分析其与精液常规、精子膜功能、精子受精功能和精子成熟障碍指标的关系。结果:头部具有透明质酸结合位点的精子,顶体完整性[(95.4±3.9)%]、线粒体膜高电位率[(97.8±2.1)%]明显高于原始活动精子[(68.8±6.2)%和(72.8±7.4)%,P均0.01];精子-透明质酸结合率与多项精液常规参数呈弱相关性(r=0.195~0.268,P均0.05),与诱发顶体反应率和精子正常形态率呈正相关(r=0.666、0.417,P均0.01),与精子核蛋白不成熟度、DNA碎片率和过量残留胞质率呈负相关(r=-0.266、-0.308、-0.218,P0.01、0.01、0.05)。结论:头部存在透明质酸受体的精子,其质膜结构、受精潜能、成熟度俱佳;用于检测精子透明质酸受体的精子-透明质酸结合试验,是独立的精子多重质量评价新指标。     

基于三维激光扫描技术的颌面三维测量与直接测量法的比较

目的：设计基于三维激光扫描技术的颌面三维测量系统,并与直接测量法进行比较。方法通过三维激光扫描仪扫描28例志愿者正面、左侧面和右侧面的3幅图像,对图像进行坏点去除、补洞等前期处理,拼接经过前期处理的3幅图像并生成整体三维模型,利用自行开发的软件3D-FACE VIEWER进行颌面部直线、曲线、角度及比例的测量,并将测量结果与直接测量法得到的结果进行对比。结果三维扫描测量法的结果与直接测量法的结果相差不大。在56项点距测量中,49项测量误差不超过2~3 mm;在17项弧长测量中,7项测量误差不超过2~3 mm。结论基于三维激光扫描技术的颌面三维测量系统是一种良好可行的方法,三维测量系统3D-FACE VIEWER可以较准确地完成颌面的测量,具有一定的临床应用价值。