周转神经损伤后碱性成纤维细胞生长因子mRNA表达的实验研究

目的 研究大鼠坐骨神经钳夹伤后碱性成纤维细胞生长因子（ｂＦＧＦ）ｍＲＮＡ表达的变化规律。方法 １０２只Ｗｉｓｔａ大鼠随机分为正常组６只,对照及实验组各４８只,用地钳钳夹坐骨神经,于伤后４小时,１,３,７,１０,１４,２１,２８天采用逆转录－聚合酶链反应技术（ＲＴ－ＰＣＲ）结合聚丙烯酰胺凝胶电泳及ａｌｐｈａ－３２ｐ－ｄＣＴＰ放射自显影,检测大鼠坐骨神经钳夹伤局部,腰４－６背根节及相应脊髓节段ｂＦＧＦ     

核因子-κB抑制剂增强131I治疗甲状腺癌疗效的裸鼠实验研究

目的 通过小鼠活体研究,探讨核因子-κB抑制剂Bay 11-7082在131I治疗DTC中的协同作用.方法 通过裸鼠腹腔注射37 MBq131I,制备清除甲状腺裸鼠模型.将72只种植KTC-1癌细胞的清除甲状腺裸鼠随机分为4组:单独用131I组、单独用Bay 11-7082组、联合用药组和对照组.给药方法为:131I剂量37 MBq,于种植癌细胞第7周第1天腹腔注射;Bay 11-7082剂量按体质量10 mg/kg,于第7周第1、2和3天腹腔注射;联合用药组两药合用,剂量同上;对照组腹腔注射相同体积的生理盐水.成瘤后每周第7天测量癌结节大小,绘制癌结节体积变化曲线.裸鼠清除甲状腺前行99TcmO4-显像,131I清除甲状腺后和131I治疗种植瘤后3d行全身显像(单独用Bay 11-7082组未行治疗后显像).第7周第7天从每组中取6只裸鼠处死,对癌结节进行凋亡染色,计算凋亡指数(染色阳性细胞数/总细胞数×100％).用单因素方差分析和q检验进行数据比较.结果 裸鼠99TcmO4-显像可见甲状腺和胃显影;清除甲状腺时腹腔注射131I后显像可见甲状腺131I浓聚明显;131I治疗后荷裸鼠全身显像可见癌结节131I浓聚.从第8周末开始,3种治疗方案所致的癌结节体积改变差异有统计学意义(F=11.91 ～246.56,均P＜0.01),联合用药组癌结节体积明显小于单一用药组(q=3.36～ 14.99,均P＜0.01).对照组、131I治疗组、Bay 11-7082治疗组和联合用药组的凋亡指数分别为(0.28±0.15)％、(5.49±0.69)％、(6.82±0.72)％和(16.2l±1.57)％,差异有统计学意义(F =304.40,P＜0.01);其中联合用药组明显高于单独用药组(q=15.33和13.33,均P＜0.01).结论 裸鼠体内实验显示,通过Bay 11-7082对核因子-κB通路的抑制,可以增强131I治疗甲状腺癌的疗效,产生协同效应.     

Nogo-66受体单链RNA干扰对大鼠脊髓半切损伤的治疗效应

目的 观察Nngo-66受体(NgR)单链RNA干扰(siRNA)对大鼠脊髓半切损伤的治疗效应. 方法 建立大鼠脊髓半切损伤模型,脑室内注射筛选出的有效NgR siRNA,采用运动功能评分、辣根过氧化物酶(HRP)逆行示踪、HE染色等方法,评价NgR siRNA对损伤脊髓的治疗效应. 结果 神经功能评分示,从第4周起,siRNA治疗组的运功功能恢复好于等渗盐水组和空白对照组,但差异无统计学意义.HRP逆行神经示踪示,治疗组多数可于脊髓前角见较多的标记细胞,与等渗盐水组和空白对照组比较,差异有统计学意义(P<0.05).HE染色示脊髓损伤后8周,等渗盐水组和对照组的损伤区纤维排列紊乱,siRNA组部分大鼠损伤区见有较连续的纤维通过.结论NgR siRNA可促进脊髓损伤后的神经再生修复.     

大鼠骶丛根性撕脱伤模型的建立

目的 建立大鼠骶丛撕脱伤模唰并评价其有效性.方法 选用20只成年SD大鼠,雌雄不限,不打开椎板,在神经孔外撕断右侧L<,4-6>神经根,左侧为对照侧.术后观察各组大鼠的存活情况,对受试大鼠进行受试鼠行为学运动(Basso-Beattie-Bresnahan,BBB)评分.并对其进行体感诱发电位(SEP)及双侧股二头肌、小腿三头肌及胫前肌刺激电位检测,辣根过氧化酶(HRP)逆行永踪,双侧股二头肌、小腿三头肌及胫前肌称重及肌肉横截面行双侧对比研究,以评价造模效果.结果 (1)一般情况:19只大鼠存活,1只大鼠死亡,存活率为95.0%.实验大鼠BBB评分为(10.78±3.15)分,而正常大鼠为21分;(2)SEP检测:17只大鼠患肢未在双侧大脑皮层检测到SEP,造模成功率为89.5%;(3)HRP示踪:2只大鼠脊髓L4-6节段内可见阳性反应,17例阴性,造模成功率为89.5%;(4)双侧股二头肌,小腿三头肌及胫前肌称重及肌肉横截面对比,差异有统计学意义;(5)电镜检测撕脱侧肌肉出现萎缩、细胞核中间移位及出现肌卫星细胞等失神经支配征象.结论 通过大鼠椎管外撕脱L4-6造模,是建立大鼠骶丛撕脱伤模型的一种可而理想的方法.     

靶肌肉注射甲钴胺对大鼠周围神经损伤再生的作用

目的　通过靶肌肉 (腓肠肌 )注射不同剂量的甲钴胺 ,观察其对大鼠坐骨神经损伤的再生作用 ,为临床肌肉局部注射甲钴胺促进周围神经再生探讨较佳的用药剂量和方法。　方法选用健康Wistar大鼠 30只 ,左侧坐骨神经横断后即刻行外膜缝合 ,制备大鼠坐骨神经损伤模型。随机分为A、B、C三组 ,每组 1 0只 ,A组注射甲钴胺 30 0 μg kg,B组注射甲钴胺 1 0 0 μg kg ,C组注射同体积的等渗盐水 1ml,术后靶肌肉注射 1次 d。术后第 4周、第 8周每组分别提取 5只大鼠 ,以左小腿三头肌湿重、光镜和电镜坐骨神经形态学观察并图像分析作为检测指标 ,分析不同剂量甲钴胺对大鼠坐骨神经损伤的影响。　结果　术后 4周左小腿三头肌湿重 ,A组为 (1 .36 7± 0 .0 1 2 )g ,B组为 (1 .1 6 4± 0 .0 1 1 )g,C组为 (0 .95 0± 0 .0 0 9)g ,差异有显著性意义 (P <0 .0 5 ) ;术后 8周 ,A组为 (2 .2 0 5± 0 .0 1 5 )g ,B组为 (1 .6 1 1± 0 .0 1 3)g ,C组为 (1 .2 30± 0 .0 1 4 )g ,差异有显著性意义 (P <0 .0 5 )。术后 4周和 8周行光镜和电镜观察 ,图像分析坐骨神经髓鞘的横截面积、壁厚度 ,A组优于B组 ,B组优于C组。　结论　靶肌肉注射甲钴胺可以促进周围神经的再生 ,高剂量的甲钴胺可以更好地发挥其对损伤神经的营养诱导作用 ,值     

131I-抗ProGRP（31-98）单克隆抗体E-B5放射免疫显像及放射免疫治疗实验研究

目的了解131I标记的胃泌素释放肽前体（ProGRP）单克隆抗体E-B5在荷小细胞肺癌裸鼠的体内分布及放射免疫显像情况,观察131I—E—B5对荷小细胞肺癌裸鼠移植瘤的生长抑制作用。方法（1）分别建立荷小细胞肺癌、荷肺腺癌及荷大肠癌裸鼠模型。（2）自荷小细胞肺癌裸鼠尾静脉注射131I—E-B5后1,12,24,48,72和96h处死裸鼠并取各主要脏器组织,计算各时间点的每克组织百分注射剂量率（％ID／g）和肿瘤／非肿瘤放射性（T／NT）比值。（3）分别对荷小细胞肺癌、荷肺腺癌和荷大肠癌裸鼠模型进行131I—E—B5放射免疫显像,观察移植瘤的显影情况,并计算移植瘤体／对侧相同部位的放射性（T／B）比值。（4）以未经任何治疗处理组为对照,采用瘤内注射法观察比较3．7,7．4,14．8和22．2MBq131I-E—B5和Na131I对荷小细胞肺癌裸鼠移植瘤的抑制作用。采用SPSS13．0软件处理数据,行两样本均数t检验。结果（1）体内分布研究示：注射后72h,荷小细胞肺癌裸鼠移植瘤体的放射性摄取达到最高值,为（14．1±2．9）％ID／g,高于其他脏器及组织（t=4．11～8．58,P均〈0．05）,瘤体与各脏器组织的T／NT比值随时间延长而逐渐升高,72h时瘤体与肌肉组织的T／NT比值高达4．67±0．66。（2）放射免疫显像发现：注射131I—E—B5后24h荷小细胞肺癌裸鼠移植瘤部位放射性明显聚集,随时间延长放射性浓聚程度逐渐增强,72至96h时最为清晰。荷肺腺癌裸鼠移植瘤局部无明显的放射性浓聚。荷大肠癌裸鼠移植瘤模型显像结果与荷小细胞肺癌裸鼠模型显像结果类似,但其T／B比值低于荷小细胞肺癌裸鼠模型（t=4．29,P〈0．01）。注射后72h,荷小细胞肺癌、荷大肠癌及荷肺腺癌裸鼠的T／B比值分别为5．27±0．97,2．28±0．72和1．26±0．65。（3）131I—E-B5对荷小细胞肺癌移植瘤的生长抑制作用明显大于Na131I（t=2．88～17．77,P均〈0．05）。结论131I—E—B5对小细胞肺癌有较好的靶向作用,可以抑制小细胞肺癌移植的生长并破坏肿瘤组织,有望成为一种较为理想的小细胞肺癌放射免疫显像及放射免疫治疗药物,值得进一步深入研究。     

周围神经损伤后碱性成纤维细胞生长因子mRNA表达的实验研究

目的　研究大鼠坐骨神经钳夹伤后碱性成纤维细胞生长因子（ｂＦＧＦ）ｍＲＮＡ表达的变化规律。方法　１０２ 只Ｗｉｓｔａｒ 大鼠随机分为正常组６ 只，对照及实验组各４８ 只，用持针钳钳夹坐骨神经，于伤后４ 小时、１，３，７，１０，１４，２１，２８ 天采用逆转录－ 聚合酶链反应技术（ＲＴ－ ＰＣＲ） 结合聚丙烯酰胺凝胶电泳及ａｌｐｈａ－３２ｐ－ ｄＣＴＰ放射自显影，检测大鼠坐骨神经钳夹伤局部、腰４ ～６ 背根节及相应脊髓节段ｂＦＧＦｍＲＮＡ的动态变化。结果　实验组伤后４ 小时ｂＦＧＦｍＲＮＡ在损伤局部、脊髓及背根节的表达均开始增强，分别为１．４１±０．１０，１．７８±０ ．１０ 和２．１７±０．１２；伤后７ 天表达值最高，分别为２．７２±０．１４，３ ．６５±０．１７ 和３．９０ ±０．０６；伤后２８ 天表达值为１．２３ ±０．０６ ，１．４３±０ ．０５，１ ．７８±０．０３，降至正常及对照组水平（Ｐ＞ ０．０５）。结论　周围神经损伤后内源性ｂＦＧＦｍＲＮＡ表达增强有助于神经再生     

131I标记抗NRP-1单克隆抗体A6在荷瘤裸鼠体内分布和显像研究

目的制备131I标记的抗神经纤毛蛋白-1（NRP-1）单克隆抗体A6（131I-A6）,探讨其作为NRP-1靶向显像新型分子探针的可行性。方法（1）采用Iodogen法对A6进行131I标记,检测其标记率、放化纯和体外稳定性。（2）以人胶质瘤细胞株U87MG为实验细胞进行体外实验,测定131I—A6生物学活性、结合率及其与受体的亲和力。（3）将荷U87MG胶质瘤模型裸鼠采用随机抽样法分为5组,每组5只,分别于注射1．2MBq 131I-A6后24、48、72、96和120h处死,计算各脏器放射性摄取（％ID／g）、肿瘤／血液（T／B）和肿瘤／肌肉（T／M）比值。（4）取6只荷瘤裸鼠,以随机抽样法分为未阻断组和竞争阻断组,前组注射3．7MBq 131I-A6;后组注射3．7MBq 131I-A6和未标记的700仙gA6,均分别于注射后24、48、72、96和120h行SPECT／CT显像。采用两样本t检验对实验数据进行统计学分析。结果（1）131I—A6标记率为（95．46±3．34）％,放化纯〉95％;131I—A6在室温下PBS溶液中放置至96h,其放化纯仍〉85％。（2） 131I-A6与U87MG胶质瘤细胞特异性结合率1h达到最高值,为（15．80±1．30）％,在加入未标记的抗体A6时,U87MG细胞对131I-A6明显受抑制（t=2．862,P〈0．05）;与细胞表面抗原的亲和力（kd）为（1．67±0．14）nmol／L。（3）24h时131I—A6在荷瘤裸鼠血液放射性最高,为（8．00±1．42）％ID／g;其次是肝脏和肿瘤组织,分别为（7．68±1．56）和（6．00±1．24）％ID／g;脑、骨、肌肉组织放射性计数较低。给药后24h,T／B和T／M分别为0．78±0．10和3．20±0．30,随时间延长比值逐渐增高,在120h达到最高,分别为1．87±0．50和7．13±0．24。（4）体内显像示,注射 131I-A6后24h肿瘤略显影,随时间延长变清晰,120h显影为最清晰,阻断后未见肿瘤显影。结论 131I-A6的标记方法简单易行,标记率高,产物稳定性好,?     

131I-VIP-ASON在荷结肠腺癌裸鼠体内的抑瘤作用

目的探讨131I标记血管活性肠肽(VIP)受体介导的C-myc mRNA反义寡核苷酸复合物(131I-VIP-ASON)对荷瘤裸鼠结肠腺癌的抑制效果.方法以TlCl3为氧化剂,将131I标记于互补于C-mycmRNA的ASON.VIP与131 I-ASON通过多聚赖氨酸偶联形成放射性反义复合物(131 I-VIP-ASON);通过荷瘤裸鼠体内分布实验观察其在肿瘤中的浓集情况;以131I-ASON和131I标记的正义寡核苷酸(SON)、无义寡核苷酸(MON)为对照,观察131 I-VIP-ASON对活体肿瘤的治疗效果.组织切片观察肿瘤组织形态学变化,Envision免疫组织染色法观察反义治疗对C-myc癌蛋白表达的影响.结果注射后2 h,131I-VIP-ASON在肿瘤部位浓聚显著高于131 I-ASON(t=7.79,P＜0.01),肿瘤/肌肉放射性比值于4 h达到最高.7.4 MBq 131 I-VIP-ASON组能有效抑制肿瘤生长,肿瘤生长终点抑制时间为25.4 d,效果明显优于3.7 MBq组(q=42.61,P＜0.01),未见明显副作用.组织形态学和免疫组织化学染色观察示,注射131 I-VIP-ASON后,肿瘤出现大片坏死区和脂肪变性.131 I-VIP-ASON组C-myc蛋白表达得分值明显低于其他组(q=5.51,P＜0.01).结论131 I-VIP-ASON可明显抑制荷瘤裸鼠结肠腺癌肿瘤生长及癌蛋白表达,可望为肿瘤的诊断和治疗提供新的思路.     

重组人促甲状腺激素在131I治疗分化型甲状腺癌中的价值

目的 评价重组人TSH (rhTSH)介导DTC 131I清除甲状腺残余组织(简称清甲)治疗的安全性及有效性.方法 回顾性分析144例甲状腺全切或次全切术后接受131I清甲治疗的DTC患者.rhTSH替代组(Ⅰ组)72例使用rhTSH 0.9mg,1次/d,连续2d肌内注射;甲状腺激素撤退组(Ⅱ组)72例停用甲状腺素药物4～6周,2组均给予3.7 GBq 131I进行清甲.观察2组FT3、FT4、TSH和Tg的变化,同时观察患者怕冷、体质量增加、腹胀、便秘、动作迟缓、皮肤干燥、眶周水肿、骨痛等反应;根据131I全身显像结果,评价2组患者的131I清甲治疗效果,显像示甲状腺床区无放射性摄取或摄取率＜1％为一次清甲完全.数据比较行x2检验或t检验.结果 2组131I治疗前血清TSH水平均升高,Ⅰ组TSH明显高于Ⅱ组[(141.26±27.30)与(70.57±51.13) mU/L,t=2.435,P＜0.05],且Ⅰ组患者血清FT3、FT4水平无明显变化;2组131I治疗前血清Tg均升高.Ⅱ组患者发生不良反应统计:怕冷80.56％(58/72),体质量增加86.11％(62/72),便秘15.28％ (11/72),动作迟缓22.22％(16/72),皮肤干燥56.94％(41/72),骨痛2.78％ (2/72),无眼眶周围水肿者.Ⅰ组治疗安全性高,主要不良反应为:头晕恶心(2.78％,2/72),骨骼疼痛(2.78％,2/72),短暂性心动过速(1.39％,1/72).131I全身显像评价患者一次清甲完全率,Ⅰ组达70.83％ (51/72),Ⅱ组达66.67％(48/72),二者差异无统计学意义(x2 =0.58,P ＞0.05).结论 使用rhTSH能有效完成DTC131I治疗前准备,提高患者的生活质量,有利于残余甲状腺组织的清除.     

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.