多肽301LARSLKT307抑制心肌分化的细胞实验研究

目的 前期发现多肽301LARSLKT307可在动物水平显著抑制胚胎期心脏发育.该研究拟进一步在P19细胞系水平探究301 LARSLKT307抑制心肌分化的作用和机制.方法 利用PepDraw Tool、Uniprot和ProtParam在线生物信息学分析网站,对多肽301 LARSLKT307的结构、保守性和理化性质进行生物信息学分析.在荧光显微镜下观察FITC标记的多肽301 LARSLKT307进入P19细胞的情况.通过倒置显微镜观察多肽301 LARSLKT307对P19细胞分化的影响.实时定量PCR及蛋白质免疫印迹法检测多肽对心肌分化标志基因肌钙蛋白cTnT和转录因子GATA4表达的影响.实时定量PCR检测多肽对Notch1信号通路关键分子Notch1和下游Hey1及Hey2的影响.结果 生物信息学分析结果 显示多肽位于其前体蛋白Talin的301～307氨基酸位点,其分子量小、结构稳定、亲脂性高,且前体蛋白保守性高.荧光显微镜下可见FITC标记的多肽301 LARSLKT307可以穿过细胞膜进入P19细胞内.与对照组比较,多肽组细胞部分坏死,生长形态紊乱,细胞厚薄不一.实时定量PCR与蛋白质免疫印迹法检测显示,与对照组比较,多肽组的分化相关基因GATA4及cTnT的mRNA及蛋白表达量均有不同程度下降.实时定量PCR检测显示,与对照组比较,多肽组Notch1通路的相关基因Notch1、Hey1及Hey2的mRNA表达均有不同程度下降.结论 多肽301LARSLKT307可能是通过抑制Notch1通路关键因子的表达发挥抑制心肌分化的生物功能.     

体外循环下小儿肌部室间隔缺损的镶嵌治疗

目的 探讨镶嵌模式(hybrid procedure)治疗小儿肌部室间隔缺损(Mvsd)的手术方法及临床应用.方法 2006年1月至2010年6月,在体外循环心内直视手术下采用手术及封堵相结合的镶嵌技术矫治小儿Mvsd 45例,其中男20例,女25例;年龄52天～12岁;体重3～32 kg.7例为单个Mvsd,38例为多发性VSD.同时合并大血管错位(D-TGA)1例、法洛四联症(TOF)2例、肺动脉狭窄(PS)3例、动脉导管未闭(PDA)6例、房间隔缺损(ASD)6例、主动脉缩窄1例.均在心脏停跳后直视下将导引钢丝经三尖瓣孔自心脏右室面穿过VSD至左室面,直视下置入导引器,然后送入封堵器,完成Mvsd封堵.多发性VSD 38例,予自体心包片修补膜周部等较大的VSD,心内其他畸形同期完成矫治.结果 42例置入单枚封堵器(直径4～10 mm)、3例置入双枚封堵器(直径4～7 mm).手术经过顺利,术前左室射血分数(EF)均在正常范围,术后1天小于8月龄组EF均值低于正常,大于8月龄组EF正常,两者差异有统计学意义.术后常规每天给予5 mg/kg肠溶阿司匹林3～6个月.术后随访超声检查示封堵器位置无偏移,无残余分流,无二尖瓣、主动脉瓣反流、Ⅲ度传导阻滞及新发心律失常等.术后因重症感染放弃治疗1例,无远期死亡病例.结论 体外循环下镶嵌技术治疗小儿 Mvsd明显降低了围手术期并发症及病死率,简化了手术过程,降低了手术风险,是一种安全、有效的方法.

Abstract：

Objective To summarize the technique and clinical experience of hybrid procedure under cardiopulmonary bypass (CPB) in children with muscular ventricular septal defect (mVSD). Methods From January 2006 to June 2010, 45 cases of mVSD underwent hybrid procedure with CPB. mVSDs were closed with devices under direct vision in 45 cases. Of them, there were 20 males and 25 females. They ranged from 52 days to 12 years [mean (2.05 ±2.48) year] in age and from 3 to 30 kg [(11.93 ±7.70)kg] in body weight. Preoperatively, most of children were highly susceptible to respiratory tract infections. The hybrid approach was used in all patients with CPB under the guidance of transesophageal echocardiography (TEE). The diameter of mVSDs ranged from 2 to 7 mm under TEE. Of 45 cases, 40 patients had increased rates of pulmonary blood flow. 29 patients had left axis deviation and 12 cases had sinus arrhythmia on electrocardiography (ECG). 19 had other congenital heart lesions, including transposition of great arteries in 1 case, tetralogy of Fallot in 2, pulmonary artery stenosis in 3, patent ductus arteriosus in 6, atrial septal defects in 6) and aorta coactation in 1. The quantity of VSDs were from 1 to 7 (single, in 7; two, in 24 case; three, in 8 case; four, in 5 case and seven, in lease. 37 patients were combined with pulmonary hypertension in our cohort. 38 patients with another large VSD and 19 with other congenital heart lesions were required surgical repair at sometime. Results The hybrid procedures were undertaken in all 45 cases of this cohort. All cases were successful and no deaths occurred during operation. A total of 48 devices were implanted in 45 patients, including single devices in 42 cases (device size ranged from 4 to 10 mm) and two devices in 3 cases (device size ranged from 4 -7 mm). The average time on CPB was (58.28 ±20.70) min , while aortic crossclamp time was(34. 94 ± 14.75) min. In addition, the time on mechanical ventilation postoperatively ranged from 2 hours to 6 days. Compared to the older children, 20 infante aged less than 8 monhad a significant difference in cardiac function in the early postoperative period. One infant was given up treatment because of serious infection. Anather cases recovered with the use of supportive treatment, such as using vasoactive agents, digoxin, inhaling nitric oxide, diuresis, and so on. The enteric-coated aspirin was given at dose of 5 mg ? kg -1. day -1 for a period of 3 to 6 months as usual postoperatively. All patients attended follow-up at 1 week, 1 month, 3 months, 6 months, 1 year and 2 years post-procedure. No major complications were encountered during this period. All cases were no instance of migration of any of the devices, residual shunt, aortic regurgitation, atrioventricular valve dysfunction, Ⅲo atrial-ventricular conduction block, new arrhythmia, and so on. There are no death in long-term follow-up. Conclusion Hybrid procedure is safe and effective for the closure of congenital heart defects in children.     

烈性呼吸道细菌多重荧光PCR检测方法建立

目的 建立多重实时荧光PCR检测炭疽杆菌、鼠疫耶尔森菌及嗜肺军团菌的方法.方法设计分别针对炭疽杆菌、鼠疫耶尔森菌、嗜肺军团菌毒力因子基因和特异性结构基因的引物和荧光双标记探针,优化反应体系,建立2套均可同时检测3种细菌的多重实时荧光PCR方法,以及3种分别针对上述单一细菌的二重实时荧光定量PCR方法.结果构建的多重和二重实时荧光PCR方法,检测敏感性分别达到100模板拷贝每反应和20模板拷贝每反应,并且高浓度模板对低浓度模板的扩增检测干扰不明显,对阳性菌株均100％检出,对常见杆菌检测均为阴性.结论多重实时荧光定量PCR具有可以同时筛查、经济、快速、特异性强、灵敏度好等优点,在炭疽杆菌、鼠疫耶尔森菌、嗜肺军团菌的病原体检测方面有较大应用价值.     

单肺Fontan手术1例

病儿 男,4岁.全身发绀4年余.查体:发育正常,营养中等,心率110次/min,血压98/65 mmHg(1 mm Hg=0.133kPa),SpO20.68,双肺未闻及于湿罗音.     

5公斤以下婴幼儿先天性心脏病的体外循环管理

方法总结我院自2007年1月至2008年12月5kg以下133例低体重婴幼儿先心病的体外循环（ECC）管理。结果 ECC时间13～382（55.72）min,主动脉阻断时间0～135（34.07）min,术后死亡3例,与ECC无直接关系。结论合理的预充,足够的流量灌注,行常规超滤和改良超滤,重视机体各脏器的保护,是5 kg以下婴幼儿ECC管理的确切有效方法。     

缺血预处理对肝血流阻断肝切除影响的荟萃分析

目的通过荟萃(Meta)分析,探讨缺血预处理(IP)对肝血流阻断肝切除术后肝功能、并发症及住院天数的影响。方法计算机检索PUBMED,EMBASE,Cochrane图书馆和重庆维普、中国期刊网、万方数据库,依据纳入和排除标准收集相关随机对照试验(RCT),进行文献筛选、数据提取及质量评价,采用RevMan 4.2.2软件进行Meta分析。结果纳入8个RCT研究共5 1 1例患者,文献质量评价均为B级。Meta分析结果表明:肝血流阻断肝切除术后IP组的ALT峰值(加权均数差=-1 7 6.3 7;9 5%CI为-3 2 0.6 7~-3 0.0 6;P=0.0 2)及并发症发生率(比值比=0.6 4;9 5%CI为0.4 1~0.9 8;P=0.0 4)均低于对照组。但两组术中出血、手术时间、肝血流阻断时间、术后AST峰值、术后总胆红素峰值及住院天数无统计学差异。结论IP可降低肝血流阻断肝切除术后ALT峰值及并发症的发生率,但其对肝脏缺血/再灌注损伤的保护作用尚缺乏充足的证据。     

大动脉调转术纠治右室双出口(Taussig-Bing型)合并冠状动脉畸形

右室双出口(Taussig-Bing型)是指右室双出口伴肺动脉瓣下型室间隔缺损,手术矫治较困难,目前国际上采用心内隧道将室缺与肺动脉开口连接,然后大动脉做调转术(Switch方法),手术结果较理想.在大动脉调转术(ASO)中,冠状动脉移植是手术成功的关键,不同类型的冠状动脉畸形是术后早期和晚期死亡的危险因素之一.当左、右冠状动脉起源于单个左或右冠窦,尤其冠状动脉走行于主动脉壁内的或仅有单根冠状动脉时,手术风险明显增加[1].我院于2010年8月至9月采用大动脉调转术连1续治疗2例右室双出口(Taussig-Bing型)合并单根冠状动脉畸形患儿,手术效果满意,现报道如下.     

实时荧光定量RT-PCR技术快速检测H5亚型禽流感病毒

目的 建立快速检测H5亚型禽流感病毒HA基因的实时荧光定量RT-PCR方法,为禽流感病毒的监测和感染的预防控制提供科学的技术手段.方法 针对H5亚型禽流感病毒血凝素(HA)基因保守区域设计特异性引物与荧光探针;通过体外克隆技术获得阳性定量标准品;优化探针、引物浓度、反应体系与反应条件,提高检测方法的特异性、敏感性与重复性,并进行定量分析.结果 反应体系具有良好的扩增效率,80 min内可完成扩增检测工作;检测灵敏度为100拷贝/反应,通过外标准品建立的标准曲线在一条直线上;3个外标准品的变异系数(CV)分别为2.60％、2.47％、2.44％;H5亚型禽流感病毒培养液、确诊人感染高致病性禽流感患者标本为阳性扩增结果,而其它流感病毒临床标本及阴性对照均呈阴性.结论 本研究建立的实时荧光定量RT-PCR方法具有敏感、快速、重复性好的特点,适合于H5亚型禽流感病毒的快速定量检测.     

先心病伴肺高压患者NO水平及临床意义探讨

目的：探讨内皮舒张因子（ＥＤＲＦ）在先心病肺动脉高压中的作用。方法：报道先心病伴肺高压虱和无肺高压先心病患者手术前后一氧化氮（ＮＯ）水平的对比观察情况。同时设正常对照组。结果；先心病肺高压组ＮＯ水平明显高于无肺高压和正常组。无肺高压和正常组无明显差异，肺高压先心病患者手术后ＮＯ水平即明显下降。     

色素失禁症伴先天性心脏病一例

患儿女,3岁4个月,因颈、胸、背部广泛色素沉着3年就诊,家长诉3年前发现患儿颈、胸、背部色素沉着斑,日益增多并向四肢扩展,就诊体检时发现心脏杂音,伴哭闹后面色口周发绀,查心脏B超确诊为先天性心脏病法洛四联症,此次为心脏手术入院.