脂质体介导的IGF-1R反义寡核苷酸抗胰腺癌的实验研究

目的研究辐射促进脂质体（hpofectin）介导的胰岛素样生长因子-1受体（IGF-1R）反义寡核苷酸（ASON）抑制人胰腺癌细胞系PC-3的生长及接种裸鼠后的成瘤性。方法PC-3细胞经不同剂量^60Co照射后，绘制剂量存活曲线，选择照射剂量。用噻唑蓝（MTT）比色法比较脂质体介导的ASON（lipo—ASON）直接转染与联合照射转染对PC-3细胞的抑制效果，用流式细胞术和逆转录-聚合酶链反应（RT—PCR）方法检测两种转染方法对PC-3细胞凋亡率及IGF-1R mRNA表达的影响。裸鼠种植PC-3细胞，肿瘤直径为5mm时，比较不同条件下（肿瘤局部照射与不照射）瘤内注射IGF-1R lipo—ASON对肿瘤的抑制情况。结果Lipo—ASON联合照射对胰腺癌细胞的抑制率（86．3％）、凋亡率（53．06％）及IGF—1R mRNA水平降低程度（降低79．2％）均明显高于未照射组（51．1％、20．25％和45.6％），差异有显著性。照射后瘤内注射lipo—ASON组的肿瘤体积明显小于其他各组，空白对照组与联合照射的lipo—ASON组、Lipo—ASON组肿瘤体积差异有显著性（P〈0．01）。结论针对IGF-1R的ASON能有效抑制肿瘤生长，当与辐射联合时，可显著提高抗瘤效果。     

CMH为KBv200耐药相关中性鞘糖脂

为了研究耐药细胞株KBv200中性鞘糖脂表达的规律，采用改良的Hakomori法提取、纯化了KB及其耐药细胞株KBv200的中性鞘糖脂，行高效薄层层析，分析其含量的异同；并以糖脂合成抑制剂PPMP抑制KBv200中性鞘糖脂的合成，观察了其对KBv200耐药性的逆转作用。结果发现：KBv200的CMH和CDH表达较KB强，尤以CMH为著，PPMP能抑制KBv200细胞CMH的合成，并能逆转KBv200对长春新碱（VCR）的耐药性。说明CMH为HBv200耐药相关中性鞘糖脂，抑制耐药细胞CMH的合成可能是逆转肿瘤多药耐药的一种新方法。     

CMH为KBv_(200)耐药相关中性鞘糖脂

为了研究耐药细胞株KBv2 0 0 中性鞘糖脂表达的规律 ,采用改良的Hakomori法提取、纯化了KB及其耐药细胞株KBv2 0 0 的中性鞘糖脂 ,行高效薄层层析 ,分析其含量的异同 ;并以糖脂合成抑制剂PPMP抑制KBv2 0 0 中性鞘糖脂的合成 ,观察了其对KBv2 0 0 耐药性的逆转作用。结果发现 ,KBv2 0 0 的CMH和CDH表达较KB强 ,尤以CMH为著 ,PPMP能抑制KBv2 0 0 细胞CMH的合成 ,并能逆转KBv2 0 0 对长春新碱 (VCR)的耐药性。说明CMH为KBv2 0 0 耐药相关中性鞘糖脂 ,抑制耐药细胞CMH的合成可能是逆转肿瘤多药耐药的一种新方法。     

分割剂量电离辐射对卵巢癌细胞多药耐药的影响

目的 研究不同分割剂量电离辐射方案对卵巢癌细胞多药耐药性的影响.方法 采用卵巢癌亲本细胞SKOV3及其耐药细胞株SKVCR,分别进行假照射、单次照射(10 Gy)、常规分割照射(2 Gy ×5)和超分割照射(1 Gy×2 ×5).MTT方法检测细胞对4种化疗药物硫酸长春新碱( vincristine,VCR)、依托泊苷(etoposide,VP-16)、盐酸吡柔比星(pirarubicin,THP)和顺铂(cisplatin,DDP)敏感性的变化.Western blot检测P-gp蛋白表达量.结果 与SKOV3相比,SKVCR倍增时间增加为1.8倍,P-gp糖蛋白表达增高,4种药物对SKVCR的IC50浓度明显增加(P＜0.05).在SKOV3中,与假照组相比,单次照射后细胞对THP、DDP药敏性降低,常规分割照射后细胞对VCR、THP、VP-16药敏性降低,超分割照射使VP-16药敏性降低;在SKVCR中,与假照组相比,3个照射组对VCR、VP-16的药敏性增高,对DDP的药敏性无明显改变,单次和分割照射均可降低P-gp表达.结论 单次、分割和超分割照射在SKOV3亲本细胞中诱导多药耐药,在耐药株SKVCR中逆转对VCR、VP-16的耐药性.     

脂质体介导的IGF-1R反义寡核苷酸对胰腺癌细胞生长的抑制作用

目的　研究照射后脂质体 (lipofectin)介导的IGF 1R反义寡核苷酸 (ASON)体外抑制人胰腺癌细胞PC 3的生长情况。方法　体外6 0 Coγ射线照射后 ,观察PC 3细胞的集落形成情况 ,以了解其辐射敏感性及确定合适的辐射剂量。用MTT法比较脂质体介导的ASON直接转染与联合辐射转染对PC 3细胞的抑制效果 ,用流式细胞仪和RT PCR方法检测两种不同方法对PC 3细胞凋亡及IGF 1RmRNA表达的调控情况。结果　联合照射的脂质体介导的ASON组明显优于未照射组 (P <0 0 5 )。PC 3细胞的凋亡率及IGF 1R的mRNA水平亦均明显高于和低于未照射组 (P <0 0 5 )。结论电离辐射可促进脂质体介导的IGF 1R反义寡核苷酸转染 ,显著地降低IGF 1R的mRNA水平 ,诱导PC 3细胞的凋亡。     

PPMP调控KBv_(200)细胞株mdr1基因表达逆转其多药耐药

为研究糖脂合成酶抑制剂苯基棕榈酰胺吗啡丙醇 (DL threo 1 phenyl 2 palmitoylamino 3 morpholi no 1 propanol·HCl,PPMP)对人恶性肿瘤多药耐药细胞株KBv2 0 0 多药耐药mdr1基因的mRNA表达的调控 ,以及PPMP对细胞株KBv2 0 0 的多药耐药性的逆转作用 ,作者应用体外细胞培养技术对细胞株KBv2 0 0 进行PPMP处理 ,用RT PCR技术分析PPMP处理前后细胞mdr1的mRNA表达 ,以流式细胞仪检测PPMP处理前后KBv2 0 0 及其敏感株KB细胞内罗丹明 12 3的药物浓度变化。结果显示 5 μmol/L、15 μmol/LPPMP可部分抑制KBv2 0 0 细胞mdr1mRNA表达 ,2 5 μmol/LPPMP可完全抑制KBv2 0 0 细胞mdr1mRNA表达 ;PPMP可增加KBv2 0 0 细胞内罗丹明荧光强度 ,随着PPMP浓度的增加 ,耐药细胞内的罗丹明荧光强度逐渐增大。提示PPMP对细胞株KBv2 0 0 的多药耐药基因mdr1有抑制作用 ,并可以逆转KBv2 0 0 的多药耐药性 ,逆转作用与PPMP浓度呈正相关     

重组人p53腺病毒在乳腺癌裸鼠体内耐药逆转作用及其对P-糖蛋白表达的影响

目的 研究重组人p53腺病毒（rAd-p53）对人乳腺癌MCF-7/ADR裸鼠移植瘤的耐药逆转作用及其对耐药相关P-糖蛋白（P-glycoprotein,P-gp）表达的影响.方法 建立人乳腺癌MCF-7/ADR裸鼠耐药模型,随机分为对照组、rAd-p53组、阿霉素组、联合组,分别给予不同治疗.观察裸鼠肿瘤体积的变化,western blot检测P-gp蛋白表达情况.结果 成瘤后rAd-p53组、阿霉素组、联合组抑瘤率分别为42.05％、49.21％、69.97％,各治疗组体积与对照组相比差异有统计学意义（P＜0.05）,且联合组显著优于rAd-p53组和阿霉素组（P＜0.05）.对照组、rAd-p53组、阿霉素组、联合组P-gp表达水平分别为（0.5964±0.0806）、（0.5506±0.0127）、（0.5641±0.0055）、（0.4652±0.0982）,联合组P-gp表达水平低于其他各组.结论 Ad-p53对人乳腺癌阿霉素耐药细胞株MCF4/ADR建立的裸鼠移植瘤具有多药耐药的逆转作用,并可下调P-gp的表达.     

辐射诱导人鼻咽癌细胞CNE1、CNE2多药耐药的研究

目的研究辐射诱导及逆转药物环孢霉素A（CsA）、干扰素a（IFNa）对鼻咽癌细胞CNE1、CNE2多药耐药表达的影响。方法通过体外辐射处理诱导鼻咽癌细胞CNEl、CNE2，流式细胞术、RT-PCR分析辐射诱导前后P-gp、MRP、TOPH、GST-π在蛋白、mRNA水平表达的改变。同时，研究CsA、IFN-α逆转耐药处理对放疗诱导的多药耐药的影响。结果辐射诱导后CNE1、CNE2细胞P-gp、MRP、GST-π在蛋白、mRNA水平表达上调，辐射后给予CsA、IFN-处理可以逆转辐射诱导的P-gp、MRP蛋白表达上调。结论辐射可以诱导鼻咽癌细胞CNE1、CNE2出现多药耐药，而CsA、IFN-可以逆转辐射诱导的多药耐药。     

pEgr-hPTEN稳定转染联合辐射诱导人胶质瘤SHG-44细胞凋亡及Bcl-2表达下调

目的 探讨辐射诱导表达载体pEgr-hPTEN体外稳定转染人胶质瘤SHG-44细胞后联合X射线照射，诱导细胞凋亡的作用及凋亡相关蛋白Bcl-2表达的变化。方法 以脂质体介导携有外源野生型PTEN基因的辐射诱导表达载体pEgr-hPTEN，体外转染SHG-44细胞，筛选稳定转染的细胞克隆并扩增培养；应用电子显微镜、流式细胞仪等方法，检测稳定转染联合X射线照射对胶质瘤细胞超微结构、细胞凋亡及Bcl-2蛋白表达等特性的影响。结果 稳定转染细胞超微结构有明显的退行性改变，可见核内染色质趋边的类似早期凋亡的改变；稳定转染联合X射线照射可诱导肿瘤细胞凋亡，5Gy以内随吸收剂量的增加，早期凋亡细胞百分数明显增加，稳定转染不同剂量照射组早期凋亡细胞百分数分别为稳定转染0Gy假照组的1．5—2．3倍、为未转染照射组的1．9—4．4倍、为未转染0Gy假照组的3．4—5．1倍；同时稳定转染细胞Bcl-2蛋白表达则呈剂量依赖性下降。结论 体外PTEN基因转染联合X射线照射可诱导肿瘤细胞凋亡明显增多，Bcl-2蛋白表达下调，具有显著的肿瘤抑制作用。     

p53腺病毒重组体转染对p53缺失肿瘤细胞辐射敏感性的影响

目的观察p53腺病毒重组体（AdCMV-p53）转染对p53缺失肿瘤细胞辐射敏感性的影响。方法用腺病毒重组体（AdCMV—p53／GFP）转染经0、0．25、0．5、1．0、1．5、2．0Gyγ射线辐射的H1299（nullp53）和PC-3（nullp53）细胞，用流式细胞分析法检测外源性p53表达，用克隆形成法检测肿瘤细胞增殖能力。结果辐射联合AdCMV-p53转染组p53阳性细胞所占比例均明显高于单纯辐射组、单纯AdCMV—p53转染组和辐射联合AdCMV—GFP转染组同种细胞p53阳性率（P〈0．05）；AdCMV—p53转染不仅明显提高肿瘤细胞辐射敏感性，而且与肿瘤细胞组织来源有关。结论p53腺病毒重组体转染对p53基因缺失肿瘤细胞低剂量辐射敏感性的增强作用与肿瘤细胞来源的组织器官和细胞类型有关。     

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.     

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.     

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）