RelB基因沉默对骨髓树突细胞生物学效应的影响

目的探讨核转录因子NFκ-B/RelB基因沉默对树突细胞(DC)生物学效应的影响。方法采用细胞因子诱导培养法体外扩增小鼠骨髓DC,并经免疫磁珠纯化,获得高纯度骨髓DC后,行RelB基因沉默,获得RNAi RelB-DC,扫描电镜和透射电镜观察细胞形态特点,并利用流式细胞术分析细胞表面分子的表达水平,同种异基因混合淋巴细胞反应(MLR)法分析RNAi RelB-DC刺激异基因T细胞增殖的能力。结果扫描电镜下RNAi RelB-DC细胞表面突起较少而短,细胞表面多皱褶;透射电镜下可见细胞内多吞噬泡样结构;流式细胞术分析显示细胞表面分子MHC-II、CD86和CD40的表达水平相对较低;MLR显示该DC刺激异基因T细胞增殖的能力相对较弱。结论RelB基因沉默DC的生物学效应表现为未成熟DC的特点,这种RNAi RelB-DC作为一种新型的致耐受DC,有望应用于免疫耐受的诱导研究。     

高迁移率族蛋白B1对大鼠脾脏树突状细胞表面共刺激分子表达的影响

目的观察高迁移率族蛋白B1(HMGB1)对树突状细胞(dendritic cells,DC)表面共刺激分子表达的影响,并对其机制进行初步探讨。方法分离正常Wistar大鼠脾脏DC后置于96孔培养板(1×10~5/孔),采用HMGB1刺激,观察HMGB1刺激与DC表面共刺激分子CD80、CD86和主要组织相容性复合物(MHC)Ⅱ表达的时间-效应关系及剂量-效应关系。结果HMGB1刺激后,DC表面共刺激分子CD80、CD86和MHCⅡ表达分别于24～72 h明显上调(P＜0．05,0．01),其中以作用48 h后DC表面共刺激分子表达上调尤为显著(P＜0．01)；0．1μg/ml、1μg/ml、10μg/ml的HMGB1刺激均可诱导DC表面共刺激分子CD80、CD86和MHCⅡ表达增强(P＜0．05,0．01),其中HMGB1的浓度在1μg/ml时,大鼠DC表面共刺激分子CD80、CD86和MHCⅡ的表达增强最明显(P＜0．01)。结论HMGB1能诱导DC表面共刺激分子表达增强,HMGB1可能是诱导DC成熟的免疫刺激信号。     

高迁移率族蛋白B1对树突细胞作用的受体机制研究

目的 观察高迁移率族蛋白B1(HMGB1)对大鼠脾脏树突细胞(DC)作用的受体机制.方法 分离正常Wistar大鼠脾脏DC,置于96孔培养板(1×105/孔),HMGB1刺激后采用激光扫描共聚焦显微镜和流式细胞术检测DC表面晚期糖基化终末产物受体(RAGE)的表达强度.同时观察抗RAGE抗体对HMGB1刺激后DC表面共刺激分子CD80、CD86和主要组织相容性复合物(MHC)Ⅱ表达的影响.结果 1μg/ml HMGB1刺激48h后,DC表面受体RAGE表达明显上调(P＜0.01);在1:50、1:100、1:200稀释度的抗RAGE多克隆抗体作用后,HMGB1刺激诱导DC表面CD80、CD86和MHC Ⅱ表达减弱(P＜0.01),其中1∶100稀释度时表达减弱最明显.结论 HMGB1能诱导DC受体RAGE表达增强,RAGE可能是参与HMGB1诱导DC成熟分化的重要受体.     

小鼠脾脏树突状细胞的分布与形态学观察

目的:认识脾脏树突状细胞的含量、分布及形态特征.方法:运用光镜、电镜观察,免疫组化标记(CD11c、CD205、CD80与CD86、I-Ab)与免疫荧光标记方法,观察小鼠脾脏树突状细胞的含量、分布与形态结构.结果:小鼠脾脏白髓与红髓中散在分布着体积较大的不规则形树突状细胞,含量以白髓边缘区最多,依次为白髓和红髓脾索.CD11c DC约占脾脏细胞总数的3%,而CD205 DC约占脾脏细胞总数的2‰.脾脏树突状细胞表面形成数个胞浆突起.胞核略长多有凹陷.胞浆低电子密度,细胞器不发达,无溶酶体和吞噬体.结论:多种树突状细胞标志物联合标记,结合光、电镜形态学观察,能够较全面反映小鼠脾脏内树突状细胞的形态分布特征,为树突状细胞研究提供了形态学依据.     

吞噬凋亡淋巴细胞的未成熟树突细胞对脂多糖介导激活的抑制作用研究

目的 探讨吞噬经体外光化学法(PUVA)处理的同种异基因淋巴细胞的未成熟树突细胞(imDC)对脂多糖(LPS)介导的树突细胞(DC)激活的抑制作用.方法 分离C57BL/6小鼠骨髓细胞,经小鼠重组白细胞介素4(rmIL-4)和重组粒-巨噬细胞集落刺激因子(rmGM-CSF)共同诱导培养制备骨髓来源的C57BL/6小鼠imDC.分离BALB/c小鼠脾淋巴细胞(SP),经PUVA处理制备成PUVA-SP.在体外将BALB/c小鼠的PUVA-SP与C57BL/6小鼠骨髓来源的imDC共同培养,获得PUVA-SP DCs.在上述imDC和PUVA-SP DCs中分别加入10ng/ml LPS刺激24h,获得LPS DCs.采用ELISA法检测上清中IL-10和IL-12的浓度,流式细胞仪检测细胞表面MHC-Ⅱ、CD40、CD86和CD80的表达水平.结果 LPS刺激后,PUVA-SP DCs表面分子CD80、CD86、CD40的表达分别为21.26％、38.50％和39.78％,远远低于imDC经LPS刺激后的表达(50.58％、66.29％、71.20％,P＜0.01).LPS刺激后PUVA-SP DCs和imDCs MHC-Ⅱ的表达分别为67.29％和68.64％,差异无统计学意义(P＞0.05).LPS刺激后,PUVA-SP DCs和imDCs分泌IL-10的水平分别为435.6±13.9pg/ml和132.6±2.8pg/ml,与刺激前相比均增高,但前者增高的水平要显著高于后者(P＜0.01).LPS刺激后,imDCs分泌高水平的IL-12,其p70和p40的表达量分别为192.1±5.9和999.8±26.9pg/ml,而PUVA-SP DCs分泌IL-12的水平分别为p7018.56±1.3pg/ml,p40 15.22±1.2pg/ml,显著低于imDCs,差异有统计学意义(P＜0.01).结论 PUVA-SP DCs虽然表达不成熟表型,但其功能不同于imDC,对LPS刺激具有抑制作用,适用于诱导体内免疫耐受.     

体外光化学疗法处理的调节性树突细胞对T细胞增殖的影响

目的 探讨体外光化学疗法处理的调节性树突细胞对T细胞增殖的影响.方法 获取人外周血单个核细胞(PBMC),利用白介素-4重组人粒细、巨噬细胞集落刺激因子诱导生成未成熟的树突细胞(imDC).分离获取人脾脏淋巴细胞(SP),经光化学疗法(PUVA)处理后获得PUVA-SP;将imDC与PUVA-SP共培养得到体外光化学治疗性树突细胞(ecpDC);将imDC与SP共培养得到SPDC;imDC加入10ng/ml的脂多糖(LPS),培养1d后得到DC.收集4组细胞,检测其表面抗体CD11c、CD83、CD86的表达.采用混合淋巴细胞培养的方法 检测吞噬了供者凋亡的脾淋巴细胞后的受者imDC对受者T细胞增殖的影响.结果 经PUVA处理后细胞早期凋亡率为91.33%.表面分子CD83、CD86在ecpDC的阳性表达率为分别为22.83%±5.26%、22.06%±4.37%,与在imDC的阳性表达率(分别为15.06%±0.59%、15.19%±1.83%)相近(P>0.05),但明显低于在DC的阳性表达率(分别为99.79%±0.36%、99.85%±0.19%),差异有统计学意义(P<0.01).吞噬了供者凋亡的脾淋巴细胞后的受者imDC细胞可明显抑制受者T细胞的增殖.结论 体外光化学疗法诱导的凋亡脾淋巴细胞可抑制树突细胞的成熟,并可抑制受者T淋巴细胞的增殖.     

树突状细胞与血液系统肿瘤

树突状细胞（DC）分前体DC、未成熟DC、移行DC和成熟DC等几个阶段。成熟DC能将抗原递呈给T淋巴细胞并与之结合,能激活纯真T淋巴细胞。CD34（＋）造血前体细胞、CD14（＋）单核细胞和一些白血病细胞在多种细胞因子作用下可分化为成熟DC。体外制备DC疫苗的方法有多种。DC与血液系统疾病密切相关。本文就上述几方面的一些进展作一综述。     

氧化高密度脂蛋白对兔树突状细胞的成熟 及趋化因子分泌的影响

目的在体外观察氧化高密度脂蛋白(oxidation-high density lipoprotein,ox-HDL)诱导兔外周血源性树突状细胞(dendritic cells,DCs)成熟情况,并观察ox-HDL对DCs趋化因子巨噬细胞炎性蛋白-1α(macro-phage inflammatory protein-1 alpha,MIP-1α)和单核细胞趋化蛋白-1(macrophage chemoattractant protein-1,MCP-1)分泌的影响情况,探讨ox-HDL对DCs成熟及趋化作用的影响。方法 12只新西兰大白兔心尖穿刺采集外周血标本,分离培养树突状细胞;同时,冻存血浆,提取HDL并氧化修饰。培养的细胞随机分为磷酸盐缓冲液组、HDL组及ox-HDL组,分别加入磷酸盐缓冲液、HDL、ox-HDL干预24 h后收集细胞,检测细胞表面CD86、主要组织相容性复合物Ⅱ(major histocompatibility complexⅡ,MHCⅡ)、CD14表达。分别于12、24、48 h取细胞培养上清液测定趋化因子MIP-1α和MCP-1。结果 ox-HDL可以上调DCs表面CD86和MHCⅡ的表达,同时下调CD14的表达,细胞的形态由不成熟往成熟转变。细胞培养上清液中MCP-1随时间和ox-HDL浓度升高而逐渐升高,而MIP-1α水平受树突状细胞是否成熟影响不大。结论 ox-HDL可以促进DCs的成熟,并增加趋化因子MCP-1的分泌,导致斑块性质的不稳定,可能与斑块的免疫反应及炎症的活化相关。     

CIK细胞的表型分析及生物学活性研究

目的　体外诱导培养细胞因子诱导的杀伤细胞 (CIK) ,并研究其表型及生物学活性。方法　从外周血分离淋巴细胞 ,经过IFN γ、CD3McAb及IL 2诱导并培养 ,获得大量的CIK细胞。FACS测定CIK表面CD3CD5 6、CD3、CD5 4、HLA DR、CD11a、CD2 8、CD86、CD80等CD分子表达情况及其百分率 ,3 H TdR检测其增殖能力 ,MTT法检测CIK对胃癌MGC 80 3细胞、肝癌Bel 74 0 2细胞的杀伤活性。结果　CIK细胞高度表达CD3、CD5 4、CD11a,中度表达CD3CD5 6、HLA DR、CD2 8CD5 4、CD2 8,不表达CD86、CD80 ,且具有较强的增殖能力及非MHC限制性杀瘤活性。结论　IFN γ、CD3McAb及IL 2三种细胞因子体外诱导单个核细胞可获得具有高增殖活性及较强杀瘤活性的CIK细胞     

失血合并闭合性骨折致小鼠脾脏树突状细胞抗原递呈功能下降

目的 研究失血合并闭合性骨折对小鼠脾脏树突状细胞（DC）抗原递呈功能的影响并初步探讨其机制。方法 致伤后24小时分离小鼠脾脏DC,检测脾脏DC在体外抗原作用下刺激脾非黏附细胞增殖的能力、细胞因子分泌功能及其表面重要分子表达的情况。结果 创伤小鼠脾脏DC体外刺激脾非黏附细胞增殖的能力下降,细胞因子分泌水平发生了变化,流式细胞分析结果显示其成熟度下降。结论 失血合并闭合性骨折可致小鼠脾脏DC的抗原递呈功能下降。     

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）