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/     

光敏剂HpD、PSD-007对小鼠骨肉瘤的光动力作用

目的观察630nm激光激发不同剂量光敏剂HpD、PSD-007对小鼠皮下移植骨肉瘤的光动力效应,并对两者进行比较。方法 C3H小鼠56只左侧腰骶部接种LM-8小鼠骨肉瘤细胞,待皮下瘤块直径约7～8mm时,随机分成3个对照组:(1)比较空白对照。(2)生理盐水加光照。(3)单纯光敏剂未光照和4个光动力治疗PDT组:HpD和PSD-007各设5mg/kg、10mg/kg两组。小鼠尾静脉注射光敏剂后6h垂直瘤区行激光照射,激光波长630nm、功率密度167mW/cm2、能量密度200J/cm2、照射时间20min、光斑直径1.5cm。实验后第7天比较各组肿瘤直径、重量、抑瘤率及病理学变化。结果光动力治疗后,各光敏剂组肿瘤表面均出现了坏死,肿瘤的体积及瘤重均显著减少,与空白对照组相比有统计学意义。结论光敏剂HpD、PSD-007光动力疗法对小鼠LM-8骨肉瘤细胞有明确的抑制作用,光动力疗法是一种很有前景的骨肿瘤辅助治疗手段。     

不同色素兔眼底对光动力治疗反应的实验研究

目的 观察不同色素兔眼底对维替泊芬(verteporfin)-光动力疗法(photodynamic therapy,PDT)的光敏反应及眼底组织损伤差别.方法 青紫蓝兔和新西兰白兔各42只,随机分为PDT治疗组、单纯激光组、单纯光敏剂组和空白对照组.其中PDT治疗组青紫蓝兔和新西兰白兔各20只,注射光敏剂10 min后接受功率密度为600 mW/cm2、波长为689 nm激光照射,照射时间15～83 s,能量密度15～50 J/cm2;单纯激光组青紫蓝兔和新西兰白兔各20只,不注射光敏剂,仅接受激光照射;单纯光敏剂组和空白对照组青紫蓝兔和新西兰白兔各1只.于PDT后第1、3、7、30和60天对兔眼行荧光素眼底血管造影术(fluorescence fundus angiography,FFA)、光镜、电镜、免疫组化检查,观察不同色素兔眼底的损伤是否存在差异.结果 PDT治疗组新西兰白兔接受能量密度25 J/cm2激光照射后1 d即出现视网膜下出血,出血量随能量增强明显增加;PDT后3～7 d,激光能量密度为20 J/cm2时新西兰白兔眼视网膜下亦可见出血,FFA检查光斑处可见明显的荧光素渗漏;PDT后30～60d,新西兰白兔视网膜下出血基本吸收,同时视网膜变薄,视网膜细胞减少,FFA检查光斑处呈片状低荧光.PDT治疗组青紫蓝兔接受能量密度30 J/cm2激光照射后1 d出现视网膜下出血,出血量随能最增强而增加;PDT后3～7 d激光能量密度25 J/cm2时受照青紫蓝兔眼视网膜下可见出血,FFA检查光斑边缘荧光素渗漏;PDT后30～60 d视网膜色素增加,FFA检查光斑区呈现斑驳状低荧光.免疫组织化学检查显示,凋亡细胞指数青紫蓝兔视网膜多于新西兰白兔,两者差异有显著意义(P＜0.05).电镜下青紫蓝兔视网膜色素上皮细胞胞质内空泡状改变较新西兰白兔明显.单纯激光组和单纯光敏剂组兔眼底未见明显改变.结论 PDT治疗后,新西兰白兔眼底的损伤以脉络膜为主,青紫蓝兔眼底的损伤以视网膜色素上皮细胞及其内视网膜为主,不同色素兔PDT治疗后眼底损伤部位存在差异.     

光动力学疗法治疗进展期食管贲门癌

目的 评价光动力学疗法联合局部注射化疗药物治疗进展期食管贲门癌的疗效.方法 应用光动力学疗法(PDT)治疗食管贲门癌148例,按治疗方法分为单纯PDT组42例和PDT联合局部化疗组106例.对每一患者均先静脉滴注光敏剂癌光啉(PSD-007)3～5 mg/kg,于用药后24、48 h分别以激光照射肿瘤局部.PDT联合局部化疗组除PDT治疗外,于每次激光照射前肿瘤局部注射化疗药物5-FU或平阳霉素.照射2次为1疗程,间隔1月重复治疗.结果 经治疗后多数患者吞咽困难、下咽痛等临床症状不同程度缓解,其中PDT联合局部化疗组吞咽困难的缓解率显著高于单纯PDT组.PDT联合局部化疗组的近期显效率显著高于单纯PDT组,而二组近期有效率则无明显差异.部分病例随访12～36个月,PDT联合局部化疗组的平均生存期较单纯PDT组显著延长.结论 PDT联合局部化疗治疗食管贲门癌的治疗方法安全有效,并能协同提高PDT的治癌效果,有临床推广应用价值.     

光动力学疗法过程中小鼠皮肤蛋白质谱变化的研究

目的 寻找皮肤对血卟啉单甲醚光动力学疗法(hematoporphyrin monomethyl ether mediated photodyrnamic therapy,HMME-PDT)应答的蛋白质分子标志物.方法 雌性BALB/c小鼠尾静脉注射血卟啉单甲醚(hematoporphyrin monomethyl ether,HMME)10 mg/kg后即刻以532 nm的激光对其皮肤进行照射,功率密度50 mW/cm2,光斑直径1 cm,照光时间20 min;同时设立单纯照光组、单纯光敏剂组以及空白对照组.术后24 h从照射局部皮肤取材,以裂解液提取皮肤蛋白,然后用蛋白质芯片(WCX2)和SELDI蛋白质芯片阅读机检测小鼠皮肤差异蛋白质的表达.结果 HMME-PDT能够明显改变照射局部皮肤蛋白质谱的表达,在质荷比为相对分子质量(Mr)4949～18 828之间,共有7个蛋白质峰的强度发生了显著改变,其中2个蛋白质表达上调,另外5个蛋白质表达下调.单独照光组在Mrl4 927处有1个蛋白质的表达下调,与PDT处理组在该点的变化一致,且与正常对照相比有显著差异.结论 PDT照射能显著改变小鼠皮肤蛋白质谱的表达,单纯照光也能引起个别蛋白的表达发生改变,并且与PDT在该点的表达趋势一致,提示这些差异蛋白质的表达与皮肤对PDT所产生的氧化应激或照光引起的光热效应的应答机制有关.     

一种新型亚苄基环戊酮类光敏剂对白念珠菌生物膜的光动力杀伤效应研究

目的:研究一种新型亚苄基环戊酮类光敏剂介导的光动力对生物膜内不同生长阶段白念珠菌的杀伤规律及生物膜胞外多糖的损伤作用,初步探讨该光敏剂对真菌生物膜的光动力效应及作用机制。方法:(1)光动力对生物膜内白念珠菌存活率的影响:实验分为PDT组、单纯光敏剂组、单纯照光组和空白对照组四组。PDT组:不同浓度光敏剂P(10,25,50和100μM)与生物膜避光孵育1 h后,532 nm激光照射,根据光照剂量分为两组:PDT1组功率密度为40 m W/cm~2,照射时间10、15和30 min;PDT2组光照时间为10 min,功率密度为40、60和80 m W/cm~2。单纯光敏剂组:只加入光敏剂,孵育1 h,不予激光照射。单纯照光组:不加入光敏剂,生物膜中加入PBS后给予激光照射。空白对照组:不加入光敏剂,不予激光照射。处理后用XTT法测定各组菌的存活率;(2)激光共聚焦显微镜观察P2-PDT对生物膜各组分的破坏:按光敏剂浓度分为低浓度组(10μM)和高浓度组(100μM),分别给予40和80 m W/cm~2的532 nm激光照射10 min,处理前后的生物膜内加入LIVE/DEAD和Con A荧光探针染色。结果:P2浓度为100μM时,40 m W/cm~2照射10、20和30 min(光剂量分别为24、36和72 J/cm~2)可使生物膜内细菌存活率分别下降40.4%、66.9%和88%;用80 m W/cm~2照射10 min(光剂量为48 J/cm~2)可使生物膜内细菌存活率下降99.2%,在10μM时也可使存活率下降92.1%。CLSM结果显示低浓度组PDT只能灭活孢子和芽管,不能杀灭菌丝态的白念珠菌;PDT后胞外多糖荧光强度显著降低,提示PDT可破坏生物膜胞外多糖。结论:P2-PDT对生物膜内的孢子、芽管和菌丝态的白念珠菌以及胞外基质中的多糖都有灭活作用;孢子和芽管对P2-PDT的敏感性比菌丝强;P2-PDT对生物膜内菌的灭活是浓度依赖和光依赖。     

不同光照剂量的光动力疗法对小鼠肝癌移植瘤增殖及局部免疫细胞的影响

目的探讨不同光照剂量的光动力疗法（PDT）对小鼠Heps肝癌移植瘤的抗肿瘤作用及局部免疫效应。方法 69只ICR小鼠皮下接种Heps肝癌细胞建立荷瘤鼠模型,随机分为对照组、光照低剂量PDT组（能量密度135 J/cm2）和光照高剂量PDT组（能量密度215～225 J/cm2）。治疗后定期测量各组肿瘤体积,计算抑瘤率。观察HE染色的瘤组织病理学改变,免疫组化法检测肿瘤组织的CD8＋细胞毒性T淋巴细胞（CD8＋CTLs）。结果 （1）光照低剂量PDT组和光照高剂量PDT组抑瘤率分别为73.9%、96.3%;光照高剂量PDT组的抑瘤效果显著高于光照低剂量PDT组（P〈0.01）。（2）PDT后2、24和72 h肿瘤组织先后出现肿瘤细胞空泡样变性、广泛核固缩及凝固性坏死;肿瘤中微血管淤血扩张、出血和管壁破坏,上述病理改变光照高剂量PDT组更为显著。（3）治疗后72 h光照低剂量PDT组和光照高剂量PDT组肿瘤中CD8＋CTLs数量均明显高于对照组（P〈0.01,P〈0.05）;而光照低剂量PDT组肿瘤中CD8＋CTLs数量又显著高于光照高剂量PDT组（P〈0.05）。结论光照高剂量PDT组的抑瘤效果显著高于光照低能剂量PDT组;PDT可引起细胞毒性T淋巴细胞在肿瘤局部浸润增加,光照低剂量PDT组对肿瘤局部CD8＋CTLs的免疫增强效应优于光照高剂量PDT组。     

竹红菌乙素脂质体对鸡冠皮肤光动力效应的初步研究

目的　观察竹红菌乙素脂质体 (HB liposome)光动力学疗法对鸡冠皮肤的光敏损伤特点。方法　成年来亨鸡 4 5只 ,随机分为对照组 9只 ,PDT组 36只。PDT组动物静脉分别注射HB liposome 0 2 5、0 5、0 75mg/kg ,给药后即刻用铜蒸气激光照射 ,功率密度分别为 5 0、1 0 0mW/cm2 ,能量密度分别为 30、6 0、1 2 0J/cm2 ,于照射后第 1、3、7、1 4、2 8天进行肉眼和光镜观察。结果　光敏剂剂量为 0 2 5、0 5mg/kg时 ,以功率密度为 1 0 0mW /cm2 的铜蒸气激光混合光照射 2 0min ,肉眼和光镜观察显示来亨鸡鸡冠的真皮乳头层毛细血管网破坏明显 ,表皮层无明显损伤 ,褪色效果明显。结论　HB liposome光动力学疗法对鲜红斑痣模型—来亨鸡鸡冠有着良好的褪色效果。     

光动力学疗法联合5-氟尿嘧啶治疗小鼠移植性实体肿瘤

目的　探讨增强光动力学疗法(PDT) 治疗恶性肿瘤疗效的有效途径。方法　80 只昆明种小白鼠于皮下接种小鼠肝癌细胞,待肿瘤长到6 ～8 mm ,随机分为先化疗后PDT、先PDT 后化疗、单纯PDT、单纯化疗和空白对照共5 组,每组16 只。化疗方法为腹腔注射5氟尿嘧啶75 μg/g ;PDT 方法为经尾静脉注射血卟啉单醚20μg/g 后3 h 以能量密度135 J/cm2 的铜蒸气激光照射肿瘤部位。结果　采用不同治疗措施的4 组小鼠瘤体缩小,而空白对照组小鼠瘤体迅速增大,各治疗组与空白对照组比较,差异均有显著意义( P< 0-05) 。各治疗组中以先化疗后PDT 组瘤体缩小尤为显著,与其他3 组比较,差异有显著意义( P< 0 .05) 。结论　PDT 和化疗均有抑瘤效应,两种方法联合运用可提高对小鼠移植瘤的疗效,而先化疗后PDT 治疗的疗效优于先PDT 后化疗。     

血卟啉单甲醚-光动力学疗法治疗实验性脉络膜新生血管

目的 观察光敏剂血卟啉单甲醚-630 nm半导体激光器光动力学疗法对实验性脉络膜新生血管的疗效及影响疗效的因素.方法 BN大鼠共21只,随机分为空白对照组?和PDT治疗组(T1～T6).氪激光光凝诱导BN大鼠建立脉络膜新生血管动物模型后14～21d,T1～13组大鼠股静脉注射血卟啉单甲醚15 mr/ks后20 min,分别以400、600、1 000 mW/cm2功率密度,波长630 nm半导体激光照射90s,即能最密度36、54、90 J/cm2;T4～T6组大鼠在给予血卟啉单甲醚30mg/kg后,以波长630 nm半导体激光分别照射60、90、150 s,即能量密度分别为36、54、90 J/cm2.眼底光斑直径皆为800～1 000μm.治疗后24 h、7 d行荧光素眼底血管造影检查和组织病理学检查,治疗后7 d行脉络膜平片的免疫组化检查.结果 荧光素眼底血管造影显示各治疗参数的CNV闭合率不同,随能量密度的增加和光敏剂剂量的增加而提高.T1、T2、T3组PDT后第24小时CNV闭合率分别为39.1%、65.2%、87.5%,PDT后第7天闭合率分别为O%、5.6%、45.5%.T4、T5、T6组PDT后第24小时CNV闭合率分别为56.5%、78.3%、90.9%,PDT后第7天闭合率分别为25.0%、68.8%、93.3%.PDT后第24小时除T1组外,其余各组均可见不同程度的CNV和脉络膜毛细血管闭合及血栓形成;T2组视网膜毛细m管和脉络膜大血管未闭合;T4、T5组部分视网膜毛细血管有附壁红细胞聚集未堵塞管腔;T3、T6组视网膜毛细血管、CNV、脉络膜毛细血管及大部分脉络膜大血管闭合.PDT后第7天各组闭塞的视网膜血管和脉络膜大血管未见完全开放.脉络膜平片的免疫组化分析显示对形成血管腔和未形成血管腔的内皮细胞均有特异性表达,用方差分析统计方法结果显示C组与T1～T6组问差异有统计意义(P<0.01).结论 血卟啉单甲醚—光动力学疗法治疗实验性脉络膜新生血管疗效确切.     

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.     

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).     

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.     

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.     

Bronchial stump fistula :treatment with covered retrievable hinged metallic stents-preliminary clinical experience

Objective To evaluate the preliminaily clinical efficacy and retrievability of a retrievable hinged covered metallic stent in the treatment of the bronchial stump fistula （BSF）. Methods Between April 2003 and March 2005, 8 patients with bronchial stump fistula after pneumonectomy or lobectomy were treated with two types （A and B） of retrievable hinged covered metallic stents. Type A stent was placed in 6 patients and type B in 2 under fluoroscopic guidance. The stent was removed with a retrieval set when BSF was healed or complications occurred. Results Stent placement in the bronchial tree was technically successful in all patients, without procedure-related complications. Immediate closure of the BSF was achieved in all patients after the procedure. Stents were removed from all patients but one. Removal of the stents was difficult in two patients due to tissue hyperplasia. Patients were followed up for 6 - 21 months. Placement of the stents remained stable in all patients except one due to severe cough. Permanent closure of BSF was achieved in 7 （87.5%） of 8 patients. Conclusion Use of a retrievable hinged covered expandable metallic stent is a simple, safe, and effective procedure for closure of the BSF. Retrieval of the stent seems to be feasible. （J Intervent Radiol, 2007, 16： 253-257）     

Prevalence and severity of hip and groin pain in sub‐elite male football: a cross‐sectional cohort study of 695 players

The purpose of this study was twofold: (a) to investigate the prevalence of hip and groin pain in sub‐elite male adult football in Denmark and (b) to explore the association between prevalence and duration of hip and groin pain in the previous season with the Copenhagen Hip and Groin Outcome Score (HAGOS) in the beginning of the new season. In total 695 respondents from 40 teams (Division 1–4) were included. Players completed in the beginning of the new season (July–Sept 2011) a self‐reported paper questionnaire on hip and/or groin pain during the previous season and HAGOS. In total 49% (95% CI: 45–52%) reported hip and/or groin pain during the previous season. Of these, 31% (95% CI: 26–36%) reported pain for >6 weeks. Players with the longest duration of pain during the previous season had the lowest HAGOS scores, when assessed at the beginning of the new season, P < 0.001. This study documents that half of sub‐elite male adult football players report pain in the hip and/or groin during a football season. The football players with the longest duration of pain in previous season displayed the lowest HAGOS scores in the beginning of the new season.