PCR-RSSO基础上HLA-Ⅰ、Ⅱ基因分型的研究

目的通过对PCR-DNA技术的分析,探讨人类白细胞抗原(HLA)基因分型方法。方法采用PCR反向序列特异性寡核苷酸(polymerase chain reaction-reverse sequence specific oligonucleotide,PCR-RSSO)杂交技术,建立改良半量扩增体系全自动HLA-I、Ⅱ等位基因分型方法,进行了635份血液标本HLA-A、B、C、DR、DQ等位基因分型,其中166份DNA同时采用序列特异性引物技术(PCR-SSP)和手工全量扩增体系PCR-RSSO技术。对全自动半量PCR-RSSO、PCR-SSP、手工PCR-RSSO 3种方法做两两比较。结果全自动半量PCR-RSSO的分型成功率为98.4%(3 124/3 175),PCR-SSP为98.8%(656/664),手工PCR-RSSO为88.3%(733/830)。经χ2检验,全自动半量PCR-RSSO与PCR-SSP的分型成功率无统计学差异,与手工PCR-RSSO有显著差异(P<0.05)。结论PCR-RSSO可识别HLA-Ⅰ,Ⅱ共706个等位基因,覆盖WHO命名委员会2000年公布的936个等位基因的75.43%;对706个HLA等位基因的分型均为中~高分辨率,有分辨纯合子等位基因的能力;易长期保存书面的实验原始资料,即杂交条;具有成本低、劳动强度低、省时和DNA消耗量少等优点。PCR-RSSO适合于造血干细胞移植和建立造血干细胞及脐带血干细胞库的组织配型。     

股骨近端纤维结构不良的手术治疗

目的探讨股骨近端纤维结构不良的手术治疗方式。方法对19例股骨近端纤维结构不良的不同手术治疗方式及术后疗效进行回顾性分析。结果1例术后3d引流管口渗出血清样物质，加强抗感染、营养支持及换药处理后愈合。19例均获随访，时间13—58个月。复查X线片见缺损修复区内有新骨生成改变，骨折处骨愈合；1例术后20个月因外伤致股骨转子下内固定物旁骨折再次手术发现肿瘤复发，行再次刮除植骨内固定术后14个月愈合；除1例未行内固定的病例外，余患者术后患肢功能均得到良好恢复，8—12个月可弃拐行走。结论股骨近端纤维结构不良应积极手术治疗，在彻底刮除病变和充分植骨的基础上，强调内固定的应用。     

Feasibility of establishing cerebral ischemia models by using aerocyst-blocking bilateral ascending pharyngeal artery of piglets Imaging assessment

BACKGROUND: The cerebral ischemia and ischemia/reperfusion animal models are used to simulate the human cerebrovascular diseases is one of the popular topics of neurological science recently. To study the pathophysiology, pathogenesis, prophylaxis and treatment of ischemic cerebrovascular diseases and to establish the ideal animal model that is the most similar to the human cerebral ischemia, are the topics that the people generally cared about. OBJECTIVE: To evaluate the effects of aerocyst-blocking bilateral ascending pharyngeal artery on the establishment of cerebral ischemia models by using digital subtraction angiography (DSA), magnetic resonance diffusion-weighted imaging (DWI) and magnetic resonance perfusion-weighted imaging (PWI). DESIGN: Repetitive measure animal experiment. SETTING: Zhongshan Hospital Affiliated to Dalian University. MATERIALS: The experiment was carried out in the Animal Laboratory (Provincial Laboratory), Zhongshan Hospital of Dalian Univeristy from January to May 2006. A total of 14 domestic piglets, of 6 months old, weighing 12–15 kg, of either gender, were selected from Animal Experimental Center, Dalian University. Multistar T.O.P digital subtraction angiography machine was provided by Siemens Company, German. METHODS: Aerocyst-blocking bilateral ascending pharyngeal artery was used to establish cerebral ischemia models. And then, Multistar T.O.P. DSA was used for imaging of cerebral vessels before blocking, during blocking and at 0.5 and 2 hours after ischemia perfusion. GE Signa 1.5 T supraconduction magnetic resonance imaging was used for DWI examination; in addition, PWI was used based on focal sites and areas. Otherwise, magnetic resonance imaging (MRI) was used to detect signal changes of T1WI and T2WI in ischemic areas. MAIN OUTCOME MEASURES: Analytic results of DSA, DWI, PWI and MRI. RESULTS: All 14 experimental piglets were involved in the final analysis. ① DSA: The blood flow of bilateral ascending pharyngeal arteries and its branch were blocked at blocking phase, which restored 0.5 and 2 hours after reperfusion. ② DWI and PWI: There were no observable abnormalities in PWI and DWI at pre-blocking. Abnormal increased signals were found on both DWI and PWI at during and post-blocking. There were reduction in ADC and rCBF and delay in rTTP at all time points except pre-blocking. ③ MRI: There were no abnormal signals observable at any time of pre- and post-blocking in T1WI and T2WI. CONCLUSION: It is feasible to establish this kind of animal experimental models, and it can simulate the ischemic state; meanwhile, the existence and extent can be showed directly by DSA, DWI, and PWI.     

电视纵隔镜和CT对非小细胞肺癌术前分期的诊断性试验的评价

目的 评价电视纵隔镜和CT对非小细胞肺癌术前分期的价值。方法 2002年9月至2003年8月28例肺癌患者术前均行CT检查，颈部纵隔镜检查术23例，胸骨旁纵隔镜检查术5例。肺癌同侧纵隔淋巴结转移N2行新辅助化疗后再手术；对测纵隔淋巴结转移N3行放化疗；阴性者剖胸探查。结果 术前怀疑或已确诊的肺癌共28例中，纵隔镜诊断纵隔淋巴结转移(阳性)20例(鳞癌11例，腺癌9例)，未见纵隔淋巴结转移8例；阳性中同侧纵隔淋巴结转移N2(16例)行新辅助化疗，后再手术；对侧纵隔淋巴结转移M(4例)行放化疗；阴性者剖胸探查，术后病理证实肺腺癌3例、肺鳞癌1例(纵隔淋巴结未见转移与纵隔镜检查结果相符)，肺炎性假瘤2例，肺结核瘤2例。28例肺癌纵隔淋巴结经CT和纵隔镜检查，分别与纵隔淋巴结清扫术后病理对照，CT(诊断标准按淋巴结≥1．0cm为阳性)诊断纵隔淋巴结转移12例，其中7例证实为转移癌，5例为淋巴结炎性反应，CT诊断纵隔淋巴结阴性16例，其中病理证实4例均为转移癌，12例为淋巴结炎性反应；纵隔镜诊断纵隔淋巴结转移20例，术后病理证实20例均为转移癌；诊断8例为淋巴结炎性反应，术后病理证实其中1例纵隔淋巴结转移。CT灵敏度63．6％，特异度70．6％，阳性预测值58．3％，阴性预测值75％；电视纵隔镜灵敏度95．2％．特异度100％，阳性预测值100％，阴性预测值87．5％。结论 电视纵隔镜对非小细胞肺癌术前分期的灵敏度、特异度、阳性预测值和阴性预测值都大大高于CT，对非小细胞肺癌的综合治疗方案的制定非常重要。     

梗阻性黄疸高危因素分析

目的 :通过探讨梗阻性黄疸临床指征 ,了解其病程中的高危因素。并及时降低并发症及死亡率 ,提高临床疗效。方法 :对我科 1993年— 2 0 0 0年收治的 2 92例梗阻性黄疸病人进行性分析 ,统计年龄、梗阻时间、性质、手术时间 ,入院 2 4小时内测尿素氮 (BUN)、白蛋白 (A)、总胆红素(TBIL)、白细胞 (WBC)、血红素 (HB)、腹水 ,分析其高危因素。结果 :年龄 >6 5岁并发症发生率为2 5 .2 0 % ,病死率 3.2 5 % ;梗阻时间超过 2 0天以上者 ,并发症 2 8.87% ,病死率 5 .15 % ;肝功能Child氏B级、C级者 ,并发症发生率为 2 9.2 0 % ,病死率为 5 .31% ,延期手术并发症发生率为 2 9.4 1% ,病死率为 4 .90 %。结论 :梗阻性黄疸病人年龄越大 ,梗阻时间越长 ,肝功能越差 ,不及时手术是各种并发症发生及病死率增高的高危因素     

血栓闭塞性脉管炎的腔内治疗

Objective To investigate the value of endovascular surgery for the treatment of thromboangitis obliterans (TAO). Methods Sixteen patients (18 limbs) with thromboangitis obliterans were treated by percutaneous transluminal plasty (PTA). Results Angiographic success on the limb basis was achieved in 15 of 18 treated limbs and the initial technical success rate was 83.34%. Doppler anklebrachial index (ABI) increased from 0.33±0.16 to 0.79±0.23 one week after PTA. The follow-up time was 2～24 months, the average time was 10.84 months. The 3-month, 12-month accumulated primary patency rate were 81.33% and 60. 23% respectively. Conclusion PTA is effective primary invasive treatment for thromboangitis obliterans yielding acceptable primary clinical success with a low complication rate and resulting in moderate long-term clinical patency and a high limb salvage rate.     

髋臼内壁内移截骨全髋置换治疗成人髋臼发育不良的有限元分析及临床意义

目的 通过计算机辅助设计有限元分析,找寻髋臼内壁截骨的合适内移范围,为临床工作提供理论依据.方法 利用SolidWorks 2005软件,建立髋臼发育不良骨盆的三维模型,模拟髋臼内壁内移截骨术式,使髋臼内壁骨从未完全陷入盆腔内保持2 mm骨性接触处开始,逐渐内移至完全陷入盆腔内7 mm处,每隔1 mm为1个实验组,分成10个实验组.每组髋臼人为划成4个象限,分别对各组假体臼-骨界面间进行计算机模拟对比力学实验分析,测量出髋臼假体-骨界面间的Mises应力及剪切应力值,找寻出应力分布较为均匀的实验组.结果 Mises应力及剪切应力均有1个象限内的值较大,通过2次统计学分析计算,得出实验结果.结论 当髋臼内壁内移至未完全陷入盆腔内1 mm处到完全陷入盆腔内1 mm处的范围内,髋臼假体-骨界面间的应力分布均匀,最佳位置在完全陷入盆腔内1 mm处.     

锁定肱骨近端接骨板治疗肱骨近端骨折

目的探讨锁定肱骨近端接骨板治疗肱骨近端骨折的临床价值：方法采用切开复位AO锁定肱骨近端接骨板(LPHP)内固定治疗肱骨近端骨折12例。结果所有患随访3～13个月，平均7．5个月。临床疗效评估：优7例，良3例，进步2例，优良率为83．3％。结论LPHP具有固定可靠、减少软组织损伤、保护血运、利于关节囊和肩袖修复、有助于骨折愈合和肩关节功能恢复的特点，值得提倡。     

Inhibitory effects of benzoate on chiral inversion and clearance of N(G)-nitro-arginine in conscious rats.

N(G)-nitro-arginine (NNA) is known to exhibit stereoselective pharmacokinetics in which N(G)-nitro-d-arginine (d-NNA) has a faster clearance rate than N(G)-nitro-l-arginine (l-NNA) in anesthetized rats, and d-NNA undergoes unidirectional chiral inversion. It was postulated that chiral inversion of d-NNA was performed in a two-step pathway by d-amino acid oxidase (DAAO) followed by an unidentified transaminase. Such chiral inversion contributes (at least partially) to the pharmacokinetic stereoselectivity of NNA. This study used the selective inhibitor of DAAO, sodium benzoate, to test the above hypothesis. An i.v. bolus injection of d-NNA (32 mg/kg) and l-NNA (16 mg/kg) in conscious rats exhibited biphasic disposition with different pharmacokinetic parameters in a stereospecific manner (approximately 5-10-fold differences). Unidirectional chiral inversion of d-NNA but not l-NNA was found from these animals. In addition to its similar inhibitory effects on the d-NNA conversion and DAAO activity in kidney homogenates, sodium benzoate completely blocked chiral inversion of d-NNA and led to a smaller stereospecific difference, reflected by a nearly 50% reduction of d-NNA clearance and a 2-fold increase in t(1/2) and area under the curve of d-NNA in benzoate-pretreated rats. The results suggest that DAAO plays an essential role in chiral inversion of d-NNA and chiral inversion contributes mostly to the pharmacokinetic stereospecificity of NNA.     

刺鼠信号蛋白对自体移植皮片中酪氨酸酶的活性影响

目的 检测刺鼠信号蛋白对自体移植皮片中酪氨酸酶的活性影响,进一步认识自体移植皮片过度色素沉着的原因。方法 建立过度色素沉着的自体移植皮片的动物模型；利用免疫组织化学方法分别检测刺鼠信号蛋白作用前后自体移植皮片中酪氨酸酶的表达,并与对照治疗组及正常皮肤相比较。结果 酪氨酸酶的表达定位于表皮基底部黑色素细胞的胞浆,在大部分组织中呈阳性表达,经刺鼠信号蛋白作用后酪氨酸酶在自体移植皮片中的表达明显减少,与在其他各对照组中的表达差异有极显著意义（P＜0.01）。结论 刺鼠信号蛋白在自体移植皮片中能竞争性拮抗α-MSH的黑色素合成,使皮片着色能力降低,从而证明皮片移植后表皮细胞中α-MSH的表达上调是皮片呈过度色素沉着的重要原因。