蒿甲醚对人鼻咽癌细胞CNE-1放射敏感性的影响及其机制研究

目的 研究蒿甲醚(artemether)对人鼻咽癌CNE-1细胞放射敏感性的影响及其作用机理。方法 用MTT法检测蒿甲醚对CNE-1细胞生长的抑制效应;克隆形成法检测蒿甲醚对CNE-1细胞放射敏感性的影响;流式细胞仪检测不同处理因素对CNE-1细胞的细胞周期的影响;Western blot分析蒿甲醚对电离辐射诱导的CNE-1细胞中的细胞周期调控蛋白Cyclin B1和Wee1表达水平的影响。结果 蒿甲醚可抑制CNE-1细胞的增殖,且药物浓度和作用时间相关;20 μmol/L蒿甲醚对CNE-1细胞有放射增敏作用,其放射增敏作用随作用时间的延长而延长,作用24 h 后增敏作用最强,放射增敏比(SER)为1.481。流式细胞仪分析表明,单纯⁶⁰Co γ射线照射后G₂/M期细胞比例上升,而G₀/G₁期细胞比例下降;药物和射线联合作用后,与单纯照射组相比,G₀/G₁期细胞比例上升(t=4.59,P<0.05),G₂/M期细胞比例下降(t=10.60,P<0.05)。蒿甲醚可降低照射后CNE-1细胞内Wee1蛋白表达水平,提高Cyclin B1蛋白表达水平。结论 低浓度蒿甲醚对CNE-1 细胞有放射增敏作用;蒿甲醚通过改变细胞周期调控蛋白Cyclin B1和 Wee1表达水平,诱导G₂/M 期阻滞而发挥放射增敏作用。     

大蒜素提高人胰腺癌BXPC3细胞放射敏感性的机制研究

目的：观察大蒜素对人胰腺癌BXPC3细胞的生长及放射敏感性的影响。方法：大蒜素联合X射线处理胰腺癌BXPC3细胞,采用MTT法检测细胞的生长和增殖;细胞流式技术测定细胞周期改变以及细胞凋亡;细胞克隆形成法观察大蒜素对辐射的增敏作用。采用RT-PCR、Western blot分析细胞中Bax、Bcl-2表达变化。结果：12、24和48 h大蒜素处理导致细胞生长抑制的半数抑制浓度（IC₅₀）分别为76.24、58.34和 43.58 μmol/L。与单纯照射组相比,照射联合应用大蒜素明显抑制胰腺癌细胞的生长（t=2.74,P<0.05）,影响细胞周期的变化,显著增加G₂/M期细胞阻滞（t=11.41,P<0.05）;细胞凋亡率明显提高（t=12.36,P<0.05）,伴有Bax表达的增高（t=4.83,P<0.05）和Bcl-2的表达下调（t=3.69,P<0.05）。 结论：大蒜素可以通过诱导细胞周期阻滞及细胞凋亡而抑制胰腺癌BXPC3细胞的生长,具有放射增敏作用,凋亡相关因子Bax、Bcl-2参与了这一过程。     

西妥昔单抗联合照射对结直肠癌CL187细胞的抑制作用及机制探讨

目的 研究西妥昔单抗(C225)联合外照射对结直肠癌CL187细胞生物学效应的影响,并探讨相关分子机制。方法 结直肠癌CL187细胞分单纯照射组和C225处理的联合照射组,受6 MV X 射线照射0、4和8 Gy后24和48 h,用四甲基偶氮唑盐(MTT)比色法测吸光度(A)值,比较两组细胞死亡率的差异。利用克隆形成实验比较两组细胞增殖能力的差异。采用流式细胞仪,检测细胞周期和细胞凋亡。蛋白免疫印迹法分析两组细胞DNA-PKcs、Ku70和Ku80蛋白表达量的变化。结果 联合照射组较单纯照射组死亡细胞比例增加(t=-6.14、-6.53,P<0.05),细胞克隆形成能力下降。C225增强了射线对细胞的杀伤作用,放射增敏比SER为1.38。联合照射组G₀/G₁期细胞阻滞增加(t=-4.64, P<0.05),细胞凋亡比例增加(t=-9.16, P<0.05),DNA修复相关蛋白DNA-PKcs、Ku70和Ku80蛋白表达量减少。结论 西妥昔单抗增强照射对结直肠癌CL187细胞的杀伤作用,可能是通过影响细胞周期、细胞凋亡和DNA损伤修复基因实现的。     

榄香烯乳对人肺腺癌A549细胞放射敏感性影响及其机制的研究

目的 探讨榄香烯乳对人肺腺癌A549细胞放射敏感性的影响及其分子机制。方法 克隆形成实验检测10、20 μg/ml榄香烯乳对人肺腺癌A549细胞的放射敏感性影响。细胞分为空白对照组、单纯照射组(4 Gy X射线照射)、单纯药物组(给予10、20 μg/ml榄香烯乳);联合照射组(给予10、20 μg/ml 榄香烯乳24 h后4 Gy X射线照射)。Western blot检测DNA-PKcs、Bcl-2及P53蛋白的表达变化,并分析DNA-PKcs与Bcl-2、P53表达之间的相关性。结果 10 μg/ml榄香烯乳的放射增敏比SER_D0、SER_Dq 为1.54±0.20和1.43±0.15;20 μg/ml榄香烯乳SER_D0、SER_Dq 为1.63±0.32及1.75±0.19。与单纯照射组相比,10、20 μg/ml榄香烯乳联合照射组细胞的DNA-PKcs蛋白表达明显减少(t=7.52、8.33, P<0.05),Bcl-2蛋白表达明显减少(t=10.74、11.33, P<0.05),P53蛋白表达明显增加(t=-9.25、-7.66P<0.05)。DNA-PKcs与P53蛋白表达显著负相关(r=-0.569,P<0.05),与 Bcl-2蛋白表达显著正相关(r=0.755, P<0.05)。结论 榄香烯乳可增加人肺腺癌A549细胞的放射敏感性,其机制与下调DNA-PKcs表达抑制DNA双链损伤修复和上调p53,下调Bcl-2表达促进细胞凋亡有关。     

黄连素对乏氧环境中鼻咽癌放射增敏作用机制的研究

目的 探究黄连素对乏氧环境中鼻咽癌细胞系CNE-2及裸鼠移植瘤的增敏效应及其分子机制。方法 四甲基偶氮唑盐比色法（MTT法）检测黄连素对CNE-2细胞的增殖抑制作用,克隆形成实验观察5 μmol/L黄连素对常氧和乏氧环境中CNE-2细胞放射敏感性的影响。黄连素乏氧下作用24 h后给予8 Gy X射线照射,流式细胞技术检测CNE-2细胞凋亡率。建立鼻咽癌CNE-2细胞裸鼠移植瘤模型,观察黄连素及单次8 Gy照射对移植瘤的抑制作用。Western blot检测乏氧条件下黄连素对HIF-1α及VEGF蛋白表达的抑制作用。 结果 黄连素显著抑制CNE-2细胞增殖,且存在时间、浓度依赖效应,24 h药物作用后半数抑制剂量IC₅₀为（14.9±2.2）μmol/L。乏氧环境下,5 μmol/L黄连素处理后的克隆形成率较单纯乏氧组下降,其增敏比（SER_D0）为1.27。在常氧环境中克隆形成率较乏氧组显著增加,常氧组和常氧加药组的SER_D0分别为1.45和1.74。5和15 μmol/L黄连素能在乏氧环境中单独发挥促凋亡作用,与对照组比较差异有统计学意义（t=5.01、9.02,P<0.05）,黄连素联合照射后,凋亡率较单纯照射组增加（t=5.31、9.91,P<0.05）。 体内实验显示,照射给药组肿瘤体积的增长显著延迟于对照组及单纯给药组,肿瘤倍增时间联合组较单纯照射组及对照组差异有统计学意义（t=2.96、14.52,P<0.05）。Western blot显示乏氧条件下HIF-1α、VEGF表达升高,黄连素作用24 h后表达显著下调。结论 黄连素对乏氧环境中的CNE-2细胞及裸鼠移植瘤有放射增敏作用,其与下调乏氧环境中HIF-1α及下游因子VEGF的表达有关。     

西妥昔单抗对人食管癌放射抗拒性细胞株KYSE-150R放射敏感性的研究

目的 观察西妥昔单抗(C225)对人食管鳞癌放射抗拒性细胞株KYSE-150R的影响。方法 分次照射建立人食管鳞癌放射抗拒性细胞株KYSE-150R;相差显微镜下观察细胞的形态学差异;G显带法进行染色体分析;克隆形成实验验证KYSE-150和KYSE-150R细胞的不同放射敏感性;采用不同浓度的C225处理KYSE-150R细胞,研究对KYSE-150R细胞放射敏感性的影响。结果 KYSE-150R细胞的群体倍增时间为(25.9±0.6)h,长于亲代细胞的KYSE-150(23.6±0.2) h(t=6.6,P＜0.01);KYSE-150R细胞的染色体数目由69.3±1.9 增加到73.7±1.2(t=-8.83,P<0.01),并且观察到有染色体畸变;KYSE-150R细胞SF₂、D₀、D_q及N 值均高于KYSE-150细胞, 加入5 μg/ml的C225后,其SF₂、D₀、D_q、N与单纯照射组KYSE-150R细胞相比有显著降低;加入C225后KYSE-150R细胞G₀/G₁、G₂/M期比例增加,S期比例减少(t=-4.478~4.308, P<0.05)。结论 C225对人食管鳞癌放射抗拒性细胞株KYSE-150R有放射增敏作用。     

香兰素衍生物BVAN08对人脑胶质瘤细胞的放射增敏作用及机制研究

目的 研究香兰素衍生物6-溴异香兰素(BVAN08)对人脑胶质瘤细胞的增殖影响、放射增敏作用和相关机制,为开发新的放射增敏抗癌药物提供实验依据。方法 采用MTT法、克隆形成率法检测BVAN08对U-251细胞增殖活性和⁶⁰Coγ射线敏感性的影响(照射剂量率为2.3 Gy/min);光学显微镜观察BVAN08作用后细胞形态学变化;透射电镜检测细胞自吞噬死亡;流式细胞术检测细胞周期和凋亡的变化;Western blot检测DNA修复蛋白DNA-PKcs的表达变化。结果 6-溴异香兰素BVAN08对U-251细胞增殖有显著抑制作用(t=1.83～3.07,P＜0.05),在10～100 μmol/L浓度范围内呈剂量和作用时间依赖性,药物作用48和72 h的IC50分别为55.3 和52.7 μmol/L。60 μmol/L BVAN08作用4 h后,U-251细胞产生明显的G₂/M期阻滞, 12 h达到峰值,G₂/M阻滞达到63.3 %,并开始同时出现细胞凋亡和自吞噬死亡。BVAN08对U-251细胞有明显的放射增敏作用,而且在照射前12 h给药的增敏效果最好,20 μmol/L浓度对2 Gy照射杀伤U-251细胞的增强比为3.14。Western blot检测表明BVAN08能显著抑制DNA-PKcs的表达。结论 香兰素衍生物BVAN08具有明显抑制人脑胶质瘤U-251细胞增殖和辐射增敏作用、并同时诱发凋亡和自吞噬死亡。BVAN08对U-251细胞的辐射增敏作用可能与G₂/M期阻滞和DNA双链断裂修复关键蛋白DNA-PKcs表达的抑制敏感性有关。     

凋亡素2配体对食管癌细胞株Eca-109放射敏感性的影响

目的 研究凋亡素2配体(Apo-2 ligand,Apo2L),又称肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor-related apoptosis inducing ligand,TRAIL)对体外培养食管癌细胞株Eca-109放射敏感性的影响。方法 MTT法检测TRAIL对Eca-109细胞的毒性;克隆形成实验检测TRAIL对Eca-109细胞的放射增敏作用;流式细胞仪(FCM)技术分析TRAIL对凋亡率及细胞周期的影响。结果 100~800 μg/ml TRAIL随浓度升高对Eca-109细胞的增殖抑制率增大,药物浓度与细胞抑制率正相关(r=0.981,P<0.01),50%抑制浓度(IC₅₀)为637 μg/ml;200 μg/ml TRAIL能增加Eca-109细胞的放射敏感性,放射增敏比为1.219;不同剂量(0~8 Gy)照射后,给药组的凋亡率均高于单照射组(F=16.97、76.65、92.86、209.66,P<0.05),给药组G₂/M期细胞比例较单照射组增加(F=9.40、84.99、87.61、2025.85,P<0.05)。结论 TRAIL可以抑制Eca-109细胞的生长,诱导细胞凋亡和G₂/M期阻滞,后者可能与TRAIL的放射增敏机制有关。     

低温等离子体联合辐射对三种癌细胞系放射增敏效应的研究

目的 探讨低温等离子体对人肝癌细胞系HepG2、非小细胞肺癌细胞系A549及人宫颈癌细胞系HeLa的放射增敏作用及其机制。方法 应用克隆形成实验观察低温等离子体对3种细胞的放射增敏作用;流式细胞仪分析3种细胞周期分布、凋亡率及活性氧含量;Western blot法检测单纯照射(R组)、单纯等离子体作用(P组)及其联合辐射(P+R组)对3种细胞中Bcl-2、Caspase-3蛋白表达的影响。结果 低温等离子体对HepG2、A549及HeLa细胞均有放射增敏作用,放射增敏比(SER_D0)分别为1.28、1.32、1.29。HepG2、A549及HeLa细胞P+R组G₂/M期比例、凋亡率及活性氧含量与R组比较均明显增高(t_G2/M期=9.52、8.24、9.53,P<0.05;t_凋亡率=10.67、38.56、6.74,P<0.05;t_{活性氧含量}=9.41、15.42、13.53,P<0.05)。HepG2和A549细胞P+R组G₂/M期比例、凋亡率及活性氧含量与P组比较均明显升高(t_G2/M期=8.75、20.37,P<0.05;t_凋亡率=8.43、9.99,P<0.05;t_{活性氧含量}=4.82、5.27,P<0.05)。3种癌细胞中P+R组Bcl-2蛋白表达较R组降低,而Caspase-3蛋白表达升高。结论 低温等离子体可提高HepG2、A549及HeLa细胞系的放射敏感性,其对HepG2及A549细胞系的放射增敏机制可能与抑制亚致死损伤修复,使细胞周期阻滞在G₂/M期,以及提高细胞内ROS水平诱导细胞凋亡有关。     

FOXO4-DRI多肽增加非小细胞肺癌细胞放射敏感性研究

目的 探讨FOXO4-DRI多肽对非小细胞肺癌（NSCLC）细胞放射敏感性的影响。方法 为检测FOXO4-DRI对NSCLC细胞的影响,将H460和A549细胞按照射与给药方式分为对照组、FOXO4-DRI组、单纯照射组、照射+FOXO4-DRI组。采用剂量率为0.99 Gy/min γ射线单次照射,在照射前10 min对H460细胞给予FOXO4-DRI 6 μmol/L,A549细胞给予FOXO4-DRI 30 μmol/L。采用CCK-8法检测照射后24、48和72 h细胞存活率;用结晶紫染色法检测细胞克隆形成数目;用伤口愈合实验检测细胞迁移程度;用流式细胞术检测细胞凋亡和细胞周期的改变。结果 与对照组相比,FOXO4-DRI组H460细胞和A549细胞存活率降低（t=1.06~50.75,P<0.05）、迁移率降低（t=33.37~139.10,P<0.05）,克隆形成数目减少（t=5.20~93.48,P<0.05）。FOXO4-DRI诱导了细胞凋亡（t=2.95~42.00,P<0.05）,导致G₀/G₁细胞周期阻滞以及G₂/M期细胞比例下降（t=3.50~31.59,P<0.05）。与照射组相比,FOXO4-DRI可进一步降低辐照后的细胞存活率（t=2.94~23.40,P<0.05）,减少辐照后克隆形成数目（t=8.43~34.00,P<0.05）和细胞迁移率（t=5.25、7.56,P<0.05）,增加细胞凋亡（t=9.20~11.52,P<0.05）。照射+FOXO4-DRI组细胞,G₂/M期细胞比例进一步下降,S期细胞增加（t=3.85~17.62,P<0.05）。结论 FOXO4-DRI多肽通过促进细胞凋亡,降低细胞增殖能力和迁移率,可以提高NSCLC细胞放射敏感性。     

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.