血卟啉单甲醚光动力学疗法对体外培养的人角质形成细胞增生的影响

目的观察人角质形成细胞株Hacat对血卟啉单甲醚(hematoporphyrinmonomethylether,HMME)的吸收特性及HMME-PDT对角质形成细胞增生的影响。方法对数生长期的Hacat细胞分别与浓度为0、10、20和40μg/ml的HMME孵育1h;同时将另一实验组的Hacat细胞与10μg/ml的HMME分别孵育0和30min、1和2h,分别以流式细胞仪检测孵育时间和孵育浓度对Hacat细胞吸收HMME的影响;在此基础上,将Hacat细胞分别与0、2·5、5、10、20和40μg/ml的HMME孵育2h,用532nm的倍频Nd∶YAG激光以20mW/cm2的功率密度分别照射0、2·5、5、10、20min,同时设立完全空白对照组和单纯光敏剂组,24h后以MTT法检测Hacat细胞的存活率。结果Hacat细胞对HMME吸收量随着孵育浓度的增高和孵育时间的增加而增多。单独照光或以高达40μg/ml的HMME孵育对Hacat无明显杀伤(P>0·05)。在激光功率密度相同的情况下,HMME-PDT对Hacat细胞的杀伤呈孵育浓度和激光能量密度相关性。结论Hacat细胞对HMME吸收呈孵育浓度和孵育时间相关性,孵育浓度及激光能量密度是影响HMME-PDT杀伤效应非常重要的影响因素,在实验范围内的HMME-PDT能对Hacat细胞产生明显的杀伤作用。     

瘢痕成纤维细胞对血卟啉单甲醚的吸收特性及其光动力效应

目的探讨增生性瘢痕成纤维细胞（HSF）对光敏剂血卟啉单甲醚（HMME）的吸收特性和光动力疗法（PDT）的杀伤效应。方法取原代培养的HSF,经流式细胞仪（FCM）检测HMME浓度（0～40μg/ml）和孵育时间（0～120 min）对HSF细胞吸收HMME的影响;MTT法检测HMME-PDT对HSF细胞存活率的影响。结果在一定HMME浓度范围内（0～40μg/ml）,HSF细胞对HMME的吸收量随HMME浓度和孵育时间的增加而增大。实验范围内的HMME-PDT对HSF细胞的杀伤效应与HMME浓度和激光能量密度正相关。结论 HSF对HMME的吸收随HMME浓度和孵育时间的增加而增大,HMME-PDT处理HSF时,光敏剂HMME浓度和激光能量密度是影响细胞存活率的重要因素。     

HMME介导的光动力疗法诱导人肝癌细胞SMMC-7721凋亡的实验研究

目的:探讨光动力疗法对肝癌细胞SMMC-7721凋亡的影响。方法:以血卟啉单甲醚(HMME)为光敏剂,波长635nm激光为激发光源的光动力疗法作用于肝癌SMMC-7721细胞。细胞分别与浓度5、10、20、30μg/ml的光敏剂孵育4h后,用不同能量密度的激光照射。MTT法检测HMME的暗毒性以及光动力疗法作用24h后的细胞活性。Hoechst细胞     

竹红菌乙素-PDT与血卟啉衍生物-PDT对人肺癌细胞杀伤效应的比较研究

目的　探讨竹红菌素 光动力疗法 (HB PDT)对肺癌细胞的杀伤效应 ,并与第一代光敏剂血卟啉衍生物 (HpD)进行比较。方法　以不同浓度的竹红菌乙素 (HB)及HpD孵育细胞 ,然后分别在铜蒸汽激光混合光饱和光剂量条件下进行照光处理 ,照光后置于 37℃、5 %CO2 的孵箱中继续孵育 2 4h ,用MTT法分别测定不同浓度下的细胞存活率 ,绘制杀伤曲线并拟合曲线方程 ,根据方程求出不同光敏剂对细胞的半数杀伤浓度 (IC50 )。结果　HB PDT对肺癌细胞株具有很强的杀伤效应 ,其IC50 为 33 82ng/ml,而HpD对肺癌细胞株的IC50 为 1316 88ng/ml,二者比较差异有统计学意义 (P <0 0 1)。结论　HB PDT对肺癌细胞株的杀伤效应明显优于HpD PDT ,是一种对肿瘤细胞杀伤所需浓度范围小、杀伤强度高的新一代光敏剂。     

血卟啉单甲醚-PDT对骨髓基质细胞及K562白血病细胞杀伤效应的初步研究

目的:对比研究血卟啉单甲醚(HMME)-PDT对骨髓基质细胞及白血病K562细胞的杀伤效应。材料与方法:实验选用白血病K562细胞株及原代培养骨髓基质细胞,光敏剂为血卟啉单甲醚(HMME),KTP激光(532nm)作为照射光源,MTT法检测细胞杀伤效应。结果:在照光剂量为10J/cm2、HMME浓度为10μg/m     

血啉甲醚对体外培养的类风湿关节炎滑膜细胞光动力杀伤作用的初步研究

探讨血啉甲醚（hematoporphyrin monomethyl ether,HMME)对体外培养的类风湿关节炎患者滑膜细胞的光动力杀伤作用。方法：用四唑盐化色法（MTT）检测了不同能量的铜蒸气激光,不同浓度的HMME对体外培养的类风湿关节炎滑膜细胞的杀伤效应,并与血卟啉衍生物（hematoporphyrin derivative,HpD)进行比较。结果：（1）单纯照光及单加HMME,HpD对细胞存活率均无明显抑制作用。（2）各种浓度的HMME在不同剂理的激光作用下对细胞均有杀伤作用,浓度越高其杀伤作用越明显;HMME浓度一定,随着照光剂量的增大杀伤作用增强;不同浓度HMME介导的光动力杀伤作用所能产生的最大抑制率不同,此时所需的光剂量也不同。（3）HMME介导的光动力效应对滑膜细胞的杀伤作用受照光前孵育时间的影响。（4）相同浓度的HMME介导的光动力杀伤效应较HpD更强。结论：HMME介导的光动力效应对滑膜细胞的杀伤作用受光敏剂的浓度及照光能量密度的影响,且明显强于HpD,临床应用于滑膜切除术可能具有更大的优势。     

血卟啉单甲醚-光动力学疗法对鼠黑素瘤细胞杀伤的实验研究

目的:探讨应用国产光敏剂血卟啉单甲醚(HMME)对鼠皮肤黑素瘤B16F10细胞株行光动力学疗法(photodynamictherapy,PDT)的效应和机制。材料与方法:采用HMME作光敏剂,对B16F10细胞用633nm波长的He-Ne激光机作光源(功率密度5mW/cm2),以不同浓度的HMME经不同光剂量照射后,MTT法测定PDT     

亚苄基环戊酮介导的光动力对多重耐药铜绿假单胞菌的体外杀伤效应

目的探讨新型光敏剂亚苄基环戊酮化合物P3介导的光动力对多重耐药铜绿假单胞菌的体外杀伤效应。方法实验对象为铜绿假单胞标准菌(ATCC27853)1株和临床多重耐药菌(PA1、PA2、PA3)3株。(1)检测实验菌株与光敏剂P3的结合特性:以荧光光谱检测法检测孵育时间和孵育浓度对实验菌株与光敏剂P3结合的影响,先将4株铜绿假单胞菌与10μM光敏剂分别孵育不同时间(5、15、30、60、120和150 min),根据前期的检测结果再选择不同浓度(2. 5、5、10、25和50μM)的光敏剂P3孵育30 min。(2)观察光敏剂P3介导的PDT对铜绿假单胞菌的体外光动力抗菌效应,即PDT组(B组),按照不同浓度的光敏剂P3分为4组分别为2. 5μM(B1组)、5μM(B2组)、10μM(B3组)和25μM(B4组),药物与4种菌株的孵育时间30 min后,进行PDT处理,激光波长532 nm,功率密度40 mW/cm2,照射时间600 s,同时设立3个对照组(A组):空白对照组(A1组)、单纯照光组(A2组)和单纯光敏剂组(A3组),用稀释平板法培养24 h进行菌落计数。结果孵育时间5～30 min时,四株铜绿假单胞菌与光敏剂P3的结合量随孵育时间延长而逐渐增加;30 min后趋于饱和。浓度梯度实验结果显示,四株铜绿假单胞菌与光敏剂P3的结合量呈孵育浓度剂量依赖性增加。在相同孵育浓度和相同孵育时间条件下,四株铜绿假单胞菌与光敏剂P3的结合量未见显著差异。光敏剂P3对四株铜绿假单胞菌的PDT杀伤作用随着光敏剂P3浓度增高逐渐增强,当光敏剂P3浓度为25μM时,PDT对4株铜绿假单胞菌株均达到有效杀伤,即活性下降均4Log;光敏剂P3对铜绿假单胞标准菌和临床耐药菌的PDT杀伤效应比较,差异无统计学意义(P0. 05);单纯照光和单纯光敏剂对细菌的存活无影响。结论光敏剂P3介导的PDT对铜绿假单胞菌有良好的体外杀伤作用,其作用不受细菌耐药性的影响。     

基于新型双光子光敏剂光动力治疗小鼠肿瘤的初步研究

目的:初步探讨双光子光敏剂光动力治疗的可能性。方法:(1)PDT对肿瘤细胞的杀伤:将浓度为10μg/ml的光敏剂B2与Hela细胞共孵育4h后,应用波长为457nm激光照射,功率密度20mW/cm2,照射时间为10min,处理12h后用MTT方法检测细胞死亡率。(2)PDT对血管组织靶向损伤:小鼠尾静脉注射光敏剂B2,剂量为17.5mg/kg,即刻用457nm的激光照射小鼠耳部血管,功率密度100mW/     

新型光敏剂Ru化合物介导的光动力对铜绿假单胞菌的体外杀伤效应

目的观察新型光敏剂Ru化合物介导的光动力(Ru complex mediated Photodynamic Therapy,Ru-PDT)对5株离体铜绿假单胞菌(Pseudomonas aeruginosa)的杀伤作用,并初步研究PDT杀菌的作用位点。方法分离培养两株标准菌株ATCC 27853、PAO1(即ATCC 15692)和3株临床耐药菌株,制备成~108CFU/ml的细菌悬液。实验分为4组:PDT组、单纯光敏剂组、单纯照光组和空白对照组。(1)PDT组:不同浓度Ru光敏剂(终浓度为0.1、0.25、1.0、2.5、10和25μM)与细菌悬液混合后避光孵育30 min,用457 nm激光照射,照射时间10 min,功率密度为40 mW/cm2。(2)单纯光敏剂组:50μM光敏剂Ru化合物与细菌悬液避光混合,孵育30 min,不予激光照射。(3)单纯照光组:同浓度细菌悬液与PBS混合后,采用波长457 nm激光照射,照射时间10 min,功率密度为40 mW/cm2。(4)空白对照组:将细菌悬液与PBS混合后,不加入光敏剂,不予激光照射。将各组处理后的细菌悬液收集,10倍递增连续稀释后涂板培养24 h进行菌落计数。将PDT处理前后的细菌收集固定,制备超薄切片,通过透射电镜观察PDT作用后菌体的形态学变化。结果在本实验的光敏剂浓度范围内,PDT对铜绿假单胞菌的杀伤作用呈光敏剂浓度依赖性,以10μM光敏剂Ru,PDT处理5株菌均能达到有效杀菌。单纯照光或光敏剂孵育对细菌存活无影响。电镜观察发现PDT处理组细菌膜结构破坏严重,细菌内容物外溢。结论 Ru-PDT能对多重耐药铜绿假单胞菌有很好的体外杀菌作用;临床耐药菌的膜结构是其产生杀菌作用的重要位点。     

Knee injury and Osteoarthritis Outcome Score (KOOS) - validation of a Swedish version

The Knee injury and Osteoarthritis Outcome Score (KOOS) is a self-administered instrument measuring outcome after knee injury at impairment, disability, and handicap level in five subscales. Reliability, validity, and responsiveness of a Swedish version was assessed in 142 patients who underwent arthroscopy because of injury to the menisci, anterior cruciate ligament, or cartilage of the knee. The clinimetric properties were found to be good and comparable to the American version of the KOOS. Comparison to the Short Form-36 and the Lysholm knee scoring scale revealed expected correlations and construct validity. Item by item, symptoms and functional limitations were compared between diagnostic groups. High responsiveness was found three months after arthroscopic partial meniscectomy for all subscales but Activities of Daily Living.     

Endovascular management of carotid-cavernous fistulas

Objective To investigate endovascular treatment of traumatic direct carotid-cavernous fistulas （CCF） and their complications such as pseudoaneurysms. Methods： Over a five-year period, 22 patients with traumatic direct CCFs were treated endovascularly in our institution. Thirteen patients were treated once with the result of CCF occluded, 8 twice and 1 three times. Treatment modalities included balloon occlusion of the CCF, sacrifice of the ipsilateral internal carotid artery with detachable balloon, coll embolization of the cavernous sinus and secondary pseudoaneurysms, and covered-stem management of the pseudoaneurysms. Results All the direct CCFs were successfully managed endovascularly. Four patients developed a pseudoaneurysm after the occlusion of the CCF with an incidence of pseudoaneurysm formation of 18.2% （4/22）. A total number of 8 patients experienced permanent occlusion of the ICA with a rate of ICA occlusion reaching 36.4% （8/22）. Followed up through telephone consultation from 6 months to 5 years, all did well with no recurrence of CCF symptoms and signs. Conclusion Traumatic direct CCFs can be successfully managed with endovascular means. The pseudoaneurysms secondary to the occlusion of the CCFs can be occluded with stent-assisted coiling and implantation of covered stents.     

The acutely limping child

Acute limping may be the result of multiple pathologies in children. The differential diagnosis varies based on the age of the child. Irrespective of age, the initial imaging work-up includes AP and frog leg radiographs of the pelvis and ultrasound; MRI may sometimes be helpful. In children less than 3 years, infections and trauma are most frequent. MRI is the imaging modality of choice when osteomyelitis is clinically suspected. Between the ages of 3 and 10 years, transient synovitis of the hip and Legg-Calvé-Perthes disease are main considerations but infection, inflammation and focal bony lesions are also considered. In children over 10 years, slipped capital femoral epiphysis also is considered.     

Do Balance Board Training Programs Reduce the Risk of Ankle Sprains in Athletes?

Introduction Ankle sprains are the most common musculo-skeletal injury that occurs in athletes,particularly in sports that require jumping and landing on one foot such as soccer,and basketball(1-4).These injuries often result in significant time loss from participation,long-term disability,and have a major impact on health care costs and resources(5-8).     

High Intensity Interval Training:New Insights

KEY POINTS ·High-intensity interval training(HIT)is characterized by repeated sessions of relatively brief，intermittent exercise.often performed with an“a11 out”effort or at an intensity close to that which elicits peak oxygen uptake(i.e.，≥90%of VO2 peak).     

Developmental validation of the PowerPlex(®) ESI 16 and PowerPlex(®) ESI 17 Systems: STR multiplexes for the new European standard

In response to the ENFSI and EDNAP groups’ call for new STR multiplexes for Europe, Promega^® developed a suite of four new DNA profiling kits. This paper describes the developmental validation study performed on the PowerPlex^® ESI 16 (European Standard Investigator 16) and the PowerPlex^® ESI 17 Systems. The PowerPlex^® ESI 16 System combines the 11 loci compatible with the UK National DNA Database^®, contained within the AmpFlSTR^® SGM Plus^® PCR Amplification Kit, with five additional loci: D2S441, D10S1248, D22S1045, D1S1656 and D12S391. The multiplex was designed to reduce the amplicon size of the loci found in the AmpFlSTR^® SGM Plus^® kit. This design facilitates increased robustness and amplification success for the loci used in the national DNA databases created in many countries, when analyzing degraded DNA samples. The PowerPlex^® ESI 17 System amplifies the same loci as the PowerPlex^® ESI 16 System, but with the addition of a primer pair for the SE33 locus. Tests were designed to address the developmental validation guidelines issued by the Scientific Working Group on DNA Analysis Methods (SWGDAM), and those of the DNA Advisory Board (DAB). Samples processed include DNA mixtures, PCR reactions spiked with inhibitors, a sensitivity series, and 306 United Kingdom donor samples to determine concordance with data generated with the AmpFlSTR^® SGM Plus^® kit. Allele frequencies from 242 white Caucasian samples collected in the United Kingdom are also presented. The PowerPlex^® ESI 16 and ESI 17 Systems are robust and sensitive tools, suitable for the analysis of forensic DNA samples. Full profiles were routinely observed with 62.5 pg of a fully heterozygous single source DNA template. This high level of sensitivity was found to impact on mixture analyses, where 54–86% of unique minor contributor alleles were routinely observed in a 1:19 mixture ratio. Improved sensitivity combined with the robustness afforded by smaller amplicons has substantially improved the quantity of data obtained from degraded samples, and the improved chemistry confers exceptional tolerance to high levels of laboratory prepared inhibitors.