羟基喜树碱对异基因大鼠心脏移植物中细胞因子mRNA的影响

为探讨异基因大鼠心脏移植物内细胞因子的动态变化，并了解羟基喜树碱（HCPT）对细胞因子表达的影响。建立大鼠腹部心脏移植模型，分为同基因移植组（SD-SD）、异基因移植组（Wistar-SD)、异基因移植Wistar-SD加HCPT治疗组，移植后1、3、5、7、14天处死受者，切取脾脏及移植的心脏，应用RT-PCR法检测IL-2、IFN-γ、IL-4、IL-10mRNA表达。结果显示，同基因移植组术后1天可测到细胞因子表达，表达量低，术后5天不能测到上述细胞因子表达；IL-2、IFN-γ在异基因移植组术后7天达高峰，HCPT治疗组术后3天达高峰，峰值显著低于单纯异基因移植组，IL-4、IL-10在异基因移植组术后5天达高峰，HCPT治疗组术后7天达高峰，峰值显著高于单纯异基因移植组，提示HCPT治疗可使移植物存活时间延长，IL-2、IFN-γ表达量降低、IL-4、IL-10表达量升高。     

IL-17在小鼠皮肤移植受者外周血及移植物中的表达

目的 探讨小鼠皮肤移植后IL-17蛋白及mRNA在外周血及移植物中的表达.方法建立三组小鼠皮肤移植动物模型:同基因组(BALBc→BALB/c)、异基因组(C57BL/6→BALB/c),脾细胞组(C57BL/6→BALB/c),每组40只.各组分别于移植后1、3、5、7d处死10只动物,采用ELISA法检测小鼠血清内IL-17水平,RT-PCR检测移植皮肤中的IL-17 mRNA的表达情况.结果 三组小鼠皮肤移植后第1天IL-17蛋白及mRNA表达有升高趋势,但差异无统计学意义(P>0.05).术后第3、5、7天异基因组IL-17蛋白及mRNA表达明显高于同基因组、脾细胞组(P<0.01),而后两组间差异无统计学意义(P>0.05).结论 IL-7在小鼠皮肤移植后早期发生急性排斥反应时呈高表达,有望作为预测和纠正排斥反应的观察指标.受体输注供体脾细胞可明显减轻异基因皮肤移植的排斥反应.     

白藜芦醇对小鼠胰岛样细胞团的抗移植排斥作用及其IL-2、IL-1O表达的影响

目的探讨白藜芦醇(Res)对小鼠胰岛样细胞团(ICCs)的抗移植排斥作用及相关细胞因子表达的影响。方法采用BALB/c小鼠胚胎干细胞系B/c-ES在体外定向诱导形成ICCs,DIPA标记后移植至近交系昆明小鼠皮下。将小鼠随机分为环孢素A(CsA)组(阳性对照)、Res组、CsA+Res组和PBS组(阴性对照),每组6只,移植前3d开始给药(CsA及Res剂量均为50mg/kg),每天1次直至取材日。分别于ICCs移植后第5天和第10天取移植物常规病理切片,观察免疫排斥情况,部分切片行免疫组化染色,检测IL-2、IL-10表达情况。结果移植后第5天各组均出现不同程度的移植排斥反应,除不用免疫抑制剂的PBS组外,其余各组均有移植细胞存活;Res组和CsA+Res组IL-2表达率分别为17.5%及14.5%,明显低于CsA组(59.5%,P<0.05),而Res组与CsA+Res组比较差异无统计学意义(P>0.05);Res组和CsA+Res组IL-10阳性表达率(分别为23%和48%)明显高于CsA组(12.5%,P<0.05),且CsA+Res组明显高于Res组(P<0.05)。移植后第10天,仅CsA+Res组有少量移植细胞存活,其余各组未见移植细胞;CsA组IL-2和IL-10表达明显减弱,其余各组几乎不表达IL-2和IL-10。结论 Res可抑制小鼠ICCs移植后的急性排斥反应,其机制可能与下调IL-2表达及上调IL-10表达有关。     

负向免疫调节树突细胞对心脏移植大鼠Th1/Th2类细胞因子的影响

目的 探讨受体树突细胞(DC)与体外光化学法(PUVA)处理的供体脾淋巴细胞共培养后,对心脏移植受体Th1/ThZ类细胞因子及移植物存活的影响.方法 以DA大鼠为供体,LEW大鼠为受体,建立大鼠腹部异位心脏移植模型.分离正常的供体脾淋巴细胞(SP),制备经PUVA处理的供体脾淋巴细胞(PWA-SP).在体外将PWA-SP或SP与受体骨髓来源的未成熟DC共同培养,检测上述处理对受体DC分泌白介素-10(IL-10)及干扰素-γ-(IFN-γ)的影响.按受体术前1周静脉输注的细胞成分将受体大鼠随机分为3组:对照组(n=7),单纯输注PBS;SP DC组(n=8).输注负载供体SP的受体大鼠DC;PWA-SP DC组(n=8),输注负载PUVA处理的供体SP的受体大鼠DC.观察移植物的存活时间,移植术后6d检测受体血清中Th1细胞因子(IL-2、IFN-γ)及Th2细胞因子(IL-10),并取移植心脏标本作切片检查,确定排斥反应的病理分级.结果 受体DC与供体PUVA-SP共培养后,其分泌的细胞因子IL-10、IFN-γ的水平分别为75.3±3.7、24.3±1.2ng/ml,明显高于同供体SP共培养后的受体DC(分别为39.0±3.1ng/ml、12.1±0.5ng/ml,P<0.01).心脏移植术后,PUVA-SP Dc组受体大鼠血清中Th1细胞因子IL-2和INF-γ的浓度分别为199.3±8.4、9.0±1.2ng/ml,明显低于对照组(分别为295.0±3.2、61.0±3.2ng/ml,P<0.01)及SP DC组(分别为559.3±35.3、69.0±2.3ng/ml,P<0.01),而其Th2细胞因子IL-10水平(58.2±0.9ng/ml)明显高于对照组(19.0±0.6ng/ml,P<0.01)及SPDC组(20.1±1.6ng/ml,P<0.01).PUVA-SP DC组移植物排斥反应病理分级明显低于对照组,而对照组移植物排斥反应病理分级则低于SP DC组.与对照组(6.7±0.3d)比较,PUVA-SP DC组移植物存活时间(27.3±1.3d)显著延长(P<0.01),而SP DC组移植物存活时间短于对照组(5.5±0.3d,P<0.05).结论 PUVA-SP DC具有负向免疫调节特性,能够在移植物受体体内诱导Th2免疫偏移,下调移植物排斥反应.     

巴利昔单抗对肾移植受者血清IL-2与血肌酐的影响

目的分析巴利昔单抗对肾移植受者白细胞介素2（IL-2）的影响，评价其治疗的有效性及安全性。方法将72例肾移植受者随机分为巴利昔单抗组（n=37）和对照组（n=35）。所有病人均采用环抱素（CsA）＋霉酚酸酯（MMF）＋泼尼松（Pred）三联免疫抑制维持治疗。巴利昔单抗组：分别于术前2h和术后4d使用巴利昔单抗1．0mg／k异。检测术后2月内IL-2和血清肌酐浓度。观察急性排斥反应（AR）、不良反应、受者和移植肾存活情况。结果巴利昔单抗组IL-2和肌酐浓度明显低于对照组（P〈0．05）。共有12例病人发生AR，巴利昔单抗组3例，对照组9例（P〈0．05）。结论巴利昔单抗明显降低IL-2和血清肌酐水平和AR发生率，不良反应少，是一种强效安全的免疫抑制剂。     

重组hIL-10抗家兔皮肤移植排斥反应中血清IL-10及其抗体的检测

目的检测重组人白细胞介素-10（recombinant human interleukiu-10,rhIL-10）作用后皮肤移植家兔血清中IL-10及其抗体的变化,探讨外源性hIL-10诱生IL-10增强抑制效应和外源性hIL-10有无免疫原性,为rhIL-10的应用提供实验依据。方法家兔分为6组：①组rhIL-10低剂量,②组rhIL-10高剂量,③组环孢菌素（CsA）低剂量,④组CsA高剂量,⑤组rhIL-10＋CsA低剂量联合,⑥组生理盐水组。移植术前1d开始用药,连续用药10d,肌肉注射。术前1d、术后1d、4d、7d、14d、21d、28d分别取外周血,分离血清,以ELISA方法检测血清中IL-10及IL-10抗体水平。结果①移植家兔血清IL-10检测,术后各组结果较术前均有升高,在术后7d达到高峰,术后4d、术后7d rhIL-10低剂量及高剂量组IL-10水平高于其余各组（P〈0.05）。CsA低剂量及高剂量组IL-10水平低于rhIL-10组及联合组（P〈0.05）。②移植家兔血清抗IL-10抗体检测,术后各组检测结果较术前均无明显升高（P〉0.05）,与抗体阳性对照比较有显著差异（P〈0.05）。结论 rhIL-10抗家兔皮肤移植反应中,家兔血清IL-10含量在皮肤移植后明显升高,表明外源IL-10诱导内源性IL-10分泌,从而增强抑制排斥反应;外源IL-10注射后家兔血清中没有检测出特异性抗体,外源性rhIL-10进入机体没有免疫原性。     

T细胞疫苗联合环孢素A抑制大鼠肝移植排斥反应实验研究

目的：研究特异性T细胞疫苗接种联合环孢素A（CsA）对大鼠肝移植排斥反应的抑制作用。方法：利用LOU／CN大鼠的脾细胞免疫CHN大鼠，取后者脾细胞活化所制备的T细胞疫苗对肝移植受体的CHN大鼠进行免疫，移植时联合CsA，以大鼠肝移植后的存活时间、混合淋巴细胞培养、微量细胞毒测定对该方法抗大鼠肝移植排斥反应的作用加在证实。结果：和对照组相比较，T细胞疫苗接种联合CsA使肝移植大鼠存活期明显延长（P＜0.01），淋巴细胞转化率等排斥反应指标也具有显著性差异（P＜0.01）。结论：T细胞疫苗接种联合CsA能显著延长异品系大鼠肝移植的存活期，有明显的抗排斥反应作用。     

口服抗原对同种异体移植免疫反应的影响

将Wistar大鼠预先喂养同种异基因SD大鼠脾细胞7天，然后接受SD大鼠的皮肤移植，7天后，再接近SD大鼠的心脏移植，观察口服抗原后体外混合淋巴细胞培养（MLC）、体内迟发型超敏反应（DTH）以及心脏存活时间。口服抗原后，体外MLC及体内DTH反应均出现明显的抗原特异性降低，口服抗原组大鼠并接近供者皮肤致敏后的心脏移植存活时间达到7天，与对照组未致敏鼠的心脏移植存活时间一致；而未口服抗原组接受皮肤致敏后的心脏移植存活时间不超过2天。提示口服抗原可以使异基因抗原的特异性免疫应答降低，延长移植存活时间，阻皮肤移植物的事先致敏反应的发生，并使加速排斥反应时间明显延迟。     

调节性树突细胞对皮肤移植小鼠IL-10+调节性B细胞的影响

目的 探讨吞噬了供者凋亡淋巴细胞的未成熟树突细胞(imDC)对皮肤移植受体小鼠外周血IL-10+CD19+调节性B细胞(Breg)比例及移植物存活时间的影响.方法 以C57BL/6小鼠作为受者,BALB/c小鼠为供者,建立小鼠皮肤移植模型.分离C57BL/6小鼠骨髓细胞,经小鼠重组白细胞介素4(IL-4)和粒-巨噬细胞集落刺激因子(GM-CSF)共同诱导,制备并培养imDC.分离BALB/c小鼠脾淋巴细胞(SP),经光化学照射方法(PUVA)处理,得到供者小鼠脾淋巴细胞(PUVA-SP).在体外将PUVA-SP与C57BL/6小鼠骨髓来源的imDC共同培养,得到PUVA-SP DCs.根据受体小鼠术前接受的静脉输注成分将其随机分为4组(n=14):PUVA-SP DC组、imDC组、成熟树突细胞(maDC)组和PBS对照组.于手术前7d分别从尾静脉注入1×106个(0.2ml)PUVA-SP DC、imDC、maDC或0.2ml PBS.于移植术后观察受体小鼠的移植物存活时间、外周血IL-10+CD19+ Breg比例及IL-10的表达情况.结果 移植术后,受体小鼠外周血IL-10+CD 19+Breg细胞占CD19+B细胞的比例在PUVA-SP DC组为7.48％,明显高于imDC组(4.12％)、maDC组(3.01％)和PBS对照组(2.37％).PUVA-SP DC组小鼠血清中IL-10表达水平为58.2±0.9ng/ml,与maDC组(20.1±1.6ng/ml)、imDC组(26.2±1.3ng/ml)及PBS对照组(19.0±0.6ng/ml)比较显著升高(P＜0.01).PUVA-SP DC组移植物存活时间为62.3±2.6d,显著长于maDC组(20.7±1.9d)、imDC组(12.1±1.0d)和PBS对照组(11.0±1.3d,P＜0.01).结论 移植术前输注PUVA-SP DCs可显著延长移植物存活时间,增加受者体内IL-10的表达水平,诱导产生较多分泌IL-10的调节性B细胞.     

负载猪源性CTLA4Ig的猪胰岛细胞阻断直接识别途径在异种移植中的作用

目的：探讨负载供体源性CTLA4Ig的胰岛细胞通过阻断直接识别途径,诱导异种胰岛细胞抑制免疫耐受的可行性及有效性。方法：使用本实验小组前期构建的携带供体源性（猪）CTLA4-Ig的腺病毒载体（Adv-pCT-LA4-Ig）转染猪胰岛细胞,植入糖尿病大鼠肾包膜下,建立猪-大鼠异种胰岛移植模型,监测移植术后大鼠血糖变化及IL-4、γ-IFN的水平变化,检测移植物中pCTLA4-Ig、猪源性胰岛素（p-Insulin）表达情况。结果：1.大鼠血糖在移植术后即开始明显下降,于第3 d左右降至正常,三组大鼠血糖分别在移植术后6、7、35 d左右开始升高;2.三组大鼠术后胰岛平均存活时间为7.43±1.72 d、7.22±1.72 d、34.50±4.14 d,实验组胰岛细胞存活时间较对照组明显延长（P〈0.01）;3.细胞因子变化：（1）γ-IFN：对照组在移植术后第1 d开始轻度升高,于第3 d后开始明显升高,第7 d达到高峰,此后缓慢升高;实验组在移植术后第1～28 d波动较小,同术前相比无明显变化,在移植后第28 d开始上升,第35 d开始急剧上升;（2）IL-4：实验组在移植术后第1 d开始下降,第7 d达到谷值;对照组在移植术后第3～5 d轻微上升,在移植后第28 d开始下降至移植术前水平,第35 d开始急剧下降;4.病理学检查：实验组移植物无明显破坏,炎细胞浸润不明显;免疫组化可见大量pCTLA4-Ig、p-Insulin表达。结论：转染供体源性CTLA4-Ig的胰岛细胞通过阻断直接识别途径,可诱导异种胰岛细胞产生较长时间的免疫耐受。     

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.