自发性狼疮小鼠明胶酶A、B的表达变化及肾乐的影响

为探讨MRL/lpr自发性狼疮小鼠肾脏明胶酶A(MMP 2 )与明胶酶B(MMP 9)的表达变化 ,明确中药复方肾乐的调节作用及机制 ,将 90只 8周龄狼疮小鼠随机分为肾乐治疗组、甲基强的松龙 (MPS)治疗组与对照组 ,肾乐治疗组经饲料服用肾乐 ( 4g·kg-1·d-1) ,MPS治疗组腹腔注射甲基强的松龙 2 5mg·kg-1·d-1,共 2 0周。观察 3组小鼠的存活率、蛋白尿、肾功能与肾脏病理改变。并利用免疫组化检测明胶酶A、B的表达变化 ,明胶酶谱法检测肾脏与尿液明胶酶A、B的活性变化。结果发现 ,肾乐、MPS与对照组相比均能够延长狼疮小鼠的存活率 ( 70 %∶80 %∶5 0 % )、减少蛋白尿的出现比率 ( 4 0 %∶33.3%∶80 % )并减轻肾脏病理改变 ;与 8周龄狼疮小鼠比较 ,2 8周龄狼疮小鼠肾小球内明胶酶A、B的表达明显增加 ,肾脏与尿液中明胶酶A、B的活性均明显增加 ,与狼疮性肾炎的病理损害密切相关。提示肾乐与MPS能够明显抑制肾小球明胶酶A、B的表达及活性变化 ,这可能是肾乐减轻MRL/lpr自发性狼疮小鼠肾脏病理损害的主要机制之一。     

凝血酶受体在衰老大鼠肾组织中的表达变化特点

目的检测衰老大鼠肾小球凝血酶受体与纤维蛋白的表达变化,明确凝血酶受体活化的意义。方法选用3月、12月与24月龄Wistar大鼠,采用改良的Lendrum纤维蛋白染色法与免疫荧光法检测纤维蛋白的沉积,免疫组化方法检测肾小球凝血酶受体PAR-1与转化生长因子13（TGF-β）在肾组织中的表达。半定量PCR法检测PAR-1 mRNA的表达。结果3月、12月与24月龄大鼠肾小球内均未检测到纤维蛋白的表达。PAR1在3月龄鼠肾小球内皮、系膜与上皮细胞中广泛表达;24月龄鼠肾小球内PAR-1的表达明显减少。PAR-1 mRNA和肾小球TGF13的表达随增龄明显增加。结论衰老大鼠肾小球凝血酶的活化不伴有纤维蛋白的形成,活化的凝血酶受体可能在衰老相关肾脏结构变化中发挥重要作用。     

基于HPV16的新型伪病毒治疗MRL/lpr小鼠狼疮性肾炎的实验研究

目的 构建基于人乳头状病毒（human papilloma virus,HPV）16型的新型伪病毒,探讨对MRL/lpr小鼠狼疮性肾炎的治疗作用,为临床治疗系统性红斑狼疮提供实验依据。方法 将12只3月龄MRL/lpr狼疮小鼠随机均分为治疗组和对照组,治疗前后测定尿蛋白、血清尿素氮、肌酐及抗ds-DNA抗体滴度。结果 治疗组小鼠治疗后的尿蛋白、血清尿素氮、肌酐及抗ds-DNA抗体均下降,与治疗前有显著性差异（P〈0.05）,而对照组治疗前后无显著性差异（P〉0.05）,并且治疗组小鼠的平均生存期比对照组明显延长（P〈0.05）。结论 基于人乳头状病毒16型的新型伪病毒可显著改善MRL/lpr小鼠的免疫状况和肾脏功能,提高平均生存期。该研究结果 为临床治疗系统性红斑狼疮提供了新的启示。     

89Zr标记间充质干细胞的制备及对系统性红斑狼疮小鼠的PET监测

目的采用89Zr-oxine复合物标记间充质干细胞(MSCs), 并探讨其在系统性红斑狼疮(SLE)模型(MRL/lpr小鼠)中的PET显像情况。方法通过18F-FDG PET显像筛选SLE小鼠模型。制备89Zr-oxine用于MSCs的标记, 每106个MSCs配置89Zr-oxine 1 MBq。将89Zr-oxine标记的MSCs通过尾静脉分别注射到选出的MRL/lpr小鼠与BALB/c小鼠(均n=5)体内, 每只注射1.2×106个标记的MSCs, 注射剂量约0.2 MBq, 并于注射后2 h、6 h、1 d、3 d、7 d、10 d、14 d分别行microPET显像, 计算每克组织百分注射剂量率(%ID/g)。采用两独立样本t检验分析数据。结果成功采用89Zr-oxine标记MSCs, 标记效率约20%, 细胞活率>90%。MicroPET显像示注射后2 h时主要分布在肺、肝等部位。注射后24 h归巢至MRL/lpr小鼠(n=5)肾脏部位的MSCs数量明显增加, MSCs在MRL/lpr小鼠的肾脏摄取高于BALB/c小鼠的肾脏摄取[(8.28±1.27)与(4.33±...     

低分子肝素和尿激酶对大鼠肾小球炎症保护作用的比较

目的比较低分子量肝素和尿激酶对肾小球炎症反应的保护作用。方法3月龄雌性Wistar大鼠40只,随机分为对照组(NC组)、脂多糖(LPS)组(L组)、LPS 氨甲环酸组(LT组)、LPS 氨甲环酸 低分子量肝素组(LTH组)及LPS 氨甲环酸 尿激酶组(LTU组),每组8只。采用直接免疫荧光检测肾小球纤维蛋白沉积和CD11b阳性细胞分布情况,Western blotting检测内皮细胞细胞间黏附分子-1(ICAM-1)蛋白质表达。结果NC组大鼠肾小球内无纤维蛋白沉积和CD11b阳性细胞;ICAM-1有少量表达。与NC组比较,L组纤维蛋白沉积(9.1%±1.6%)和CD11b阳性细胞(11.2±2.1)增多(P<0.05);ICAM-1表达(0.23±0.09)明显上调(P<0.05)。与L组比较,LT组纤维蛋白沉积(23.4%±3.2%)和CD11b阳性细胞(20.4±3.5)进一步增多;ICAM-1表达(0.44±0.16)进一步上调(P<0.05)。与LT组比较,LTH组和LTU组纤维蛋白沉积(分别为11.3%±1.4%,10.6%±1.5%)和CD11b阳性细胞(分别为7.3±1.8,8.4±1.6)明显减少(P<0.05或0.01);ICAM-1表达(分别为0.19±0.10,0.18±0.09)明显下调(P<0.05),LTH、LTU两组间无统计学差异。结论低分子肝素与尿激酶均能有效减少纤维蛋白沉积,起到减轻肾小球炎症反应的作用。     

两种植物雌激素对5/6肾切除鼠残余肾组织α-平滑肌肌动蛋白沉积的影响及机制探讨

目的 探讨金雀异黄素、大豆黄素2种主要的植物雌激素对5/6肾切除鼠残余肾组织α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)沉积的影响及其机制.方法 32只鼠作5/6肾切除,将术后2周存活的27只鼠分为金雀异黄素组(9只)、大豆黄素组(10只)和对照组(8只),另有8只为假手术组.检测各组术前、术后第4、8周生化指标.观察第8 周肾组织的病理改变情况,应用免疫组化法检测肾组织中α-SMA的沉积情况,采用RT-PCR法和Western Blot法检测肾组织中转化生长因子-β1(TGF-β1)mRNA转录以及蛋白质表达情况.结果 与对照组相比,金雀异黄素组和大豆黄素组尿蛋白排泄量明显减少,血肌酐、尿素氮水平下降(P<0.01),肾小球硬化程度明显减轻,肾小球内α-SMA沉积显著减少,TGF-β1 mRNA及其蛋白质表达明显减弱(P<0.05).结论 金雀异黄素和大豆黄素可以降低5/6肾切除鼠残肾组织α-SMA的沉积,原因可能与抑制TGF-β1的表达有关.     

FVB小鼠急性缺血再灌注肾损伤中CHOP蛋白作用的探讨

目的通过对比ICR、FVB两种小鼠肾脏急性缺血再灌注后的差异,探索导致肾脏急性缺血再灌注损伤的关键致病分子,并探讨其可能机制。方法将ICR、FVB两种小鼠(雄性、8~10周龄)各随机分为模型组和正常对照组(n=12),两种小鼠的模型组采用相同的方法建立肾脏急性缺血再灌注模型(先切除右肾再夹闭左肾肾蒂45 min),再灌注24 h后收集模型组小鼠的心脏血、肾脏,检测各小鼠血浆BUN浓度,观察小鼠肾组织病理损伤的程度,Western-blot检测肾组织CHOP蛋白的表达,对照组检测同样的指标。结果(1)肾组织PAS染色可见:ICR、FVB小鼠模型组肾小管上皮细胞损伤明显重于对照组,差异有统计学意义(P<0.01),ICR小鼠模型组肾小管上皮细胞损伤又明显重于FVB小鼠模型组(P<0.01);(2)血浆BUN浓度:ICR、FVB小鼠模型组高于对照组(P<0.01),ICR小鼠模型组高于FVB小鼠模型组(P<0.01);(3)Western-blot:[CR、FVB小鼠模型组CHOP蛋白的表达量高于对照组(P<0.01),ICR小鼠模型组CHOP蛋白的表达量高于FVB小鼠模型组(P<0.05)。结论在急性肾缺血再灌注损伤模型中ICR小鼠肾小管上皮细胞损伤程度明显重于FVB小鼠,内质网应激相关蛋白CHOP表达也明显上调,提示CHOP在急性肾缺血再灌注损伤中发挥了重要的作用,FVB小鼠对急性肾缺血再灌注损伤存在抗性,机制可能与CHOP蛋白的表达有关。     

PAR1与PAR4在肿瘤转移中作用的研究进展

以往研究表明,凝血酶通过其细胞表面的受体--蛋白酶活化受体(protease-activated receptors,PARs)影响肿瘤的转移.PAR1是介导凝血酶促肿瘤转移的主要受体,表达于血管内皮细胞上的PAR1通过影响肿瘤血管新生而促进肿瘤转移.近年来研究发现,PAR1表达于多数肿瘤细胞表面,其表达水平往往预示着该细胞侵袭能力的强弱.PAR4在肿瘤转移中的作用报道较少,仅有的几篇报道提示,PAR4在血管内皮细胞中有表达且在血管发育中起作用,但其是否通过影响肿瘤血管新生而参与肿瘤转移还未见报道.同时,PAR4在肿瘤细胞上的表达仅见于肺癌组织,且其表达水平与肺癌患者的生存直接相关,提示PAR4可能是介导凝血酶影响肿瘤转移的另一重要受体.     

纤维蛋白诱导活化系膜细胞表达明胶酶A、B的机制研究

为观察纤维蛋白对系膜细胞（HMC）表达明胶酶A（MMP-2）、明胶酶B（MMP-9）的作用并探讨其活化机制,应用RT-PCR、明胶酶谱分析法分别在基因转录水平与蛋白质活性水平上检测纤维蛋白对HMC表达MMP-2、MMP-9的作用,采用纤维蛋白酶谱法与平板法检测纤维蛋白对HMC表达纤溶酶原激活物（PA）活性的影响。结果发现,纤维蛋白呈剂量与时间依赖性促进HMC MMP-2与MMP-9表达的上调,并同时促进HMCtPA与uPA的表达;应用抑肽酶阻断纤溶酶活性后能够完全阻断pro-MMP-2与pro-MMP-9的活化。提示肾脏局部沉积的纤维蛋白可以促进HMC表达与活化MMP-2、MMP-9、而MMP-2与MMP-9的活化依赖于PA/纤溶酶系统的调控。     

从细胞生物学角度深入研究活化凝血因子参与慢性进行性肾损伤的机制

传统观点认为,肾小球内凝血导致慢性进行性肾损伤的主要机制是微血栓形成引起肾小球缺血笥损伤。近年来作者采用细胞生物学方法,观察了活化凝血因子凝血酶和纤维蛋白对肾小球内皮细胞和系膜细胞的直接影响,发现凝血酶可以诱导ＧＥＣ增殖,细胞外基质降解增加以及细胞脱壁,纤维蛋白可以支持ＧＥＣ铺展,增殖,并可以诱导ＧＥＣ单层结构破坏和血样结构形成,同时可以上调胞间粘附分子－１的表达。     

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.