应用MRCP对63例肝移植术后胆道并发症的临床分析

目的评价MRCP在诊断原位肝移植术后胆道并发症中的临床应用价值。方法分析63例肝移植术后怀疑有胆道并发症患者的MRCP图像,并与手术、胆道造影、临床随访证实结果进行对照。所有病例均在高场强1.5T磁共振上进行。MRCP采用两种不同的成像方法:厚层块T2加权成像和薄层块多层T2加权成像。结果MRCP诊断移植术后胆道并发症的敏感性为95.3%(41/43),阳性预测值97.6(41/42),假阴性率为4.54%(2/44),假阳性率为2.27%(1/44)。总诊断准确率为95.2%(60/63)。MRCP作为唯一的诊断方法能为96.8%(61/63)的患者提供特异性诊断结果,仅2例患者需要ERCP和PTHC检查3.2%(2/63)。直接胆道造影作为一项治疗手段应用于22.2%(14/63)的患者中。结论MRCP是评价肝移植术后胆道并发症的有效影像学方法。     

大鼠骨髓间充质干细胞与多孔双相磷酸钙陶瓷支架体外黏附的实验研究

目的研究大鼠骨髓间充质干细胞(marrowmesenchymalstemcells,MSCs)诱导培养后在多孔双相磷酸钙(biphasiccalciumphosphate,BCP)陶瓷支架材料上的黏附和生长。方法SD大鼠MSCs经矿化诱导培养、扩增,具有成骨细胞表型后,与多孔BCP陶瓷支架及普通多孔羟基磷灰石(hydroxyapatite,HA)陶瓷支架体外复合培养,扫描电镜观察比较MSCs在两种材料支架表面的黏附数量和形态;同时以0.5、1.0、2.0、3.0和4.0×106/ml浓度细胞悬液接种于多孔BCP支架材料,检测适宜的接种浓度及单位体积支架材料可黏附MSCs数量。结果大鼠MSCs经诱导培养14d后,行矿化沉积茜素红染色、型胶原免疫细胞化学染色及碱性磷酸酶细胞化学染色,结果均为阳性。大鼠MSCs黏附于多孔BCP陶瓷上的细胞数(88.00±6.58)明显高于HA陶瓷组(39.00±3.65),两组比较差异有统计学意义(P<0.01)。当接种浓度为2.0×106/ml时,单位体积的陶瓷支架材料最多可黏附MSCs数量为1.28×107个/cm3,为细胞适宜接种浓度。结论大鼠MSCs在体外经矿化诱导培养可表达成骨细胞表型,细胞浓度为2.0×106/ml时与多孔BCP陶瓷支架材料具有良好的黏附能力。     

颈椎术后病人首次下床活动的感受及需求的调查

早期下床活动的必要性已得到大家认可,但术后早期病人常常较虚弱,且下床活动时会引发一些不适,病人也会有一些担心和顾虑,这在一定程度上会影响病人下床活动的积极性。颈椎手术本身有其特殊性,术后对颈部的稳定要求较高,因此此类病人活动有其特点。本研究以颈椎术后已下床活动的病人为研究对象,通过问卷调查,了解颈椎手术后病人首次下床的感受和相关需求,为进一步提高颈椎术后护理质量提供依据。1资料和方法1.1对象采用方便取样,选取2005年4月—2005年6月在某三甲医院因颈椎病入院、术后已下床活动的病人75例。其中男54例,女21例;年龄28岁~7…     

载氧血红蛋白纯化工艺研究

目的改善心脑血管缺氧载氧药物是一种创新药物。由于它半径比红细胞小400~1 000倍,易于通过毛细血管,给缺血组织及时供氧,迅速缓解或纠正缺氧状态,达到治疗抢救目的。血红蛋白的纯化工艺是载氧药物研制的重要工艺步骤。方法本研究建立了一套通过热敏法分离纯化人脐带血血红蛋白的工艺以及较为完善的纯化血红蛋白质量检测指标。结果与现有的纯化方式相比,热敏法操作简便,仪器设备造价低廉,纯化与病毒灭活同时进行,得到的纯化产品损失少,纯度高,各项理化指标达到国际水平。结论本工艺适用于规模制备纯化血红蛋白,为进一步研制治疗心脑血管缺氧载氧药物创造了有利条件。     

98例尿道下裂手术的体会

目的通过对尿道下裂手术的改进,进一步完善尿道下裂手术的技巧和护理。方法A组50例尿道下裂Ⅰ期成形术中,采用皮下连续内翻缝合形成新尿道,且固定于阴茎海绵体,由阴茎根部自下而上多层次,多点缝合皮瓣皮下组织至阴茎海绵体上,均无间隙和死腔。B组48例采用常规手术方式作对照。结果A组50例尿道下裂Ⅰ期成形术成功44例,B组48例成功17例,A组与B组相比成功率显著提高(P＜0．05)。结论尿道下裂Ⅰ期成形术成功率关键在于术者对术式掌握的熟练程度和术中及术后对每个细节的认真处理。     

肝移植围手术期出凝血功能障碍的防治

目的　探讨肝移植围手术期出凝血功能障碍的防治。方法　回顾性分析我院 2 0 0 2年 6月～ 2 0 0 3年 12月施行的 6 1例肝移植病例。结果　 6 1例肝移植术前肝功能ChildC级 35例 (5 7 4 % ) ,ChildB级 2 6例 (4 2 6 % ) ,ChildC组的患者术中凝血指标 (INR)的变化程度大于ChildB组 (P <0 0 5 )。与凝血有关的并发症中大出血 5例 (8 2 % ) ,肾衰 6例 (9 8% ) ,肝动脉血栓形成 5例 (8 2 % ) ,手术开展两阶段对比 ,第二阶段主要因限制了大量凝血药及血制品的使用 ,并发症明显减少。结论　掌握好不同时期、不同患者出血和血栓形成的平衡是防治肝移植围手术期出凝血功能障碍的关键     

Lethal effect of mononuclear cells derived from human umbilical cord blood differentiating into dendritic cells after in vitro induction of cytokines on neuroblastoma cells

BACKGROUND: Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human granulocyte-macrophage colony stimulating factor (rhG-MCSF) and recombinant human interleukin-4(rhIL-4) can generate a great many dendritic cells and promote the lethal effect of T cells on human neuroblastoma, but it is unclear that whether the lethal effect is associated with the most proper concentration of dendritic cells. OBJECTIVE: To investigate the lethal effect of human umbilical cord blood mononuclear cells induced in vitro by cytokines differentiating into dendritic cells on human neuroblastoma, and its best concentration range. DESIGN: Open experiment. SETTING: Department of Pediatrics, the Medical School Hospital of Qingdao University. MATERIALS: The study was carried out in the Shandong Provincial Key Laboratory (Laboratory for the Department of Pediatrics of the Medical School Hospital of Qingdao University) during September 2005 to May 2006. Human umbilical cord blood samples were taken from the healthy newborn infants of full-term normal delivery during October to November 2005 in the Medical School Hospital of Qingdao University, and were voluntarily donated by the puerperas. Main instruments: type 3111 CO2 incubator (Forma Scientific, USA), type 550 ELISA Reader (Bio-Rad, USA). Main reagents: neuroblastoma cell line SK-N-SH (Shanghai Institute of Life Science, Chinese Academy of Sciences), RPMI-1640 culture fluid and fetal bovine serum (Hyclone), rhIL-4 (Promega, USA), rhG-MCSF (Harbin Pharmaceutic Group Bioengineering Co.Ltd), rat anti-human CD1a monoclonal antibody and FITC-labeled rabbit anti-rat IgG (Xiehe Stem cell Gene Engineering Co.Ltd). METHODS: ① Human umbilical cord blood mononuclear cells obtained with attachment methods differentiated into human umbilical cord blood dendritic cells, presenting typical morphology of dendritic cells after in vitro induction by rhG-MCSF and rhIL-4. ② Different concentrations of dendritic cells[ dendritic cells: neuroblastoma cells=20∶1,50∶1,100∶1（2×108 L-1，5×108 L-1，1×109 L-1）], 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the experimental group. 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the control group. ③ Main surface marker CD1a molecules of dendritic cells were detected with indirect immunofluorescence, and the percent rate of dendritic cells was counted with ultraviolet light and expressed as the expression rate of CD1a+ cells. ④ Single effector cells and target cells were respectively set in the experimental group and control group to obtain the lethal effect. The lethal effect of dendritic cells on neuroblastoma cells was indirectly evaluated by detecting cellular survival with MTT assay. The lethal effect(%)=（1-A experimental well-A effector cell well/A target cell well）×100%.⑤The experimental data were presented as Mean ±SD, and paired t test was used. MAIN OUTCOME MEASURES: ① Morphological characters of dendritic cells in the process of induction and differentiation. ②CD1a+ cellular expression rate. ③Lethal effect of dendritic cells on neuroblastoma cells. RESULTS: ①Morphological characters of dendritic cells in the process of induction and differentiation: On the 15th day after human umbilical cord blood mononuclear cells were induced by rhG-MCSF and rhIL-4, typical morphology of dendritic cells could be seen under an inverted microscope. ②Expression rate of CD1a+ cells was （43.12±5.83）%. ③Lethal effect of dendritic cells on neuroblastoma cells: Lethal effect of dendritic cells stimulated T cells in each experimental group ( dendritic cells: neuroblastoma cells=100∶1,50∶1,20∶1 respectively) on neuroblastoma cells was significantly higher than that in control group[（31.00 ±4.41）%，（30.92±5.27）%，(33.57±5.35)%,(26.23±5.20)%, t=3.51，2.98，4.24, P < 0.01）; But the lethal effect of dendritic cells on neuroblastoma was significantly lower when their ratio was 100∶1 and 50∶1 in comparison with 20:1 （t=2.01，2.36, P < 0.05）, and no significant difference in lethal effect existed between the ratio at 100∶1 and 50∶1（t=0.06,P > 0.05）. CONCLUSION: Dendritic cells differentiated from human umbilical cord blood mononuclear cells after in vitro induction of cytokines can promote the lethal effect of T cells on neuroblastoma cells. The lethal effect is associated with the concentration of dendritic cells within some range.     

SB203580对肾缺血/再灌注损伤时细胞凋亡及p38MAPK影响的实验研究

目的　探讨不同时间及不同浓度p38MAPK特异性抑制剂SB2 0 35 80对肾缺血 /再灌注损伤过程中肾功能、细胞凋亡及 p38MAPK活性、表达量、p38MAPK底物的影响。 方法　 4 9只大鼠按缺血 /再灌注及给药时间的不同 ,随机分为 7组 ,每组7只大鼠按正交拉丁方表的顺序 ,经尾静脉注射相同体积、不同剂量的SB ,使其在大鼠体内达到不同的浓度。测定BUN和Scr;用TUNEL试剂盒检测细胞凋亡情况 ;Westernblot技术用于蛋白定性及半定量分析。结果　SB可显著减轻大鼠肾缺血 /再灌注损伤造成的Scr和BUN的升高、肾小管上皮细胞的凋亡及 p38MAPK的激活 ,但存在模型、剂量及给药时机的差异 (P<0 .0 5 ) ,缺血前 3h之前给药 ,使其体内血药浓度达到 5 μmol/L左右可取得较好的效果。 结论　SB可显著减轻大鼠肾缺血 /再灌注损伤 ,在缺血前 3h之前给药 ,同时使其血药浓度达到 5 μmol/L左右可取得最佳效果。     

D2-43病毒E蛋白在酵母细胞表面的展示

目的：在酵母细胞表面展示登革2型病毒43株(D2—43)的E基因，探索利用酵母表面展示系统建立DNA改组筛选平台的可行性。方法：通过RT-PCR扩增获得D2-43的E基因，将该基因亚克隆至T载体后，再克隆至酵母表面展示载体pYDI，于酿酒酵母EBY100中利用半乳糖进行诱导表达。表达产物采用间接免疫荧光法和FACS进行检测。结果：酵母表面展示产物可与D2-43的腹水抗体特异性地结合；在半乳糖诱导后24h，展示E蛋白的酵母细胞百分数达22．07％。结论：本研究为建立基于酵母表面展示系统的DNA改组筛选平台奠定了基础。     

UB-I粘结剂粘结面的微形态观察

目的：研究UB-Ⅰ粘结剂的粘结界面和粘结表面的微形态特点。方法：用扫描电子显微镜观察UB-Ⅰ和牙本质的粘结界面和粘结表面。结果：UB-Ⅰ粘结剂在牙本质小管内可聚合形成树脂突，在牙本质小管内的树脂突表面有树脂侧枝呈树枝状横行排列，并且和相邻的树脂突相连接，在牙本质和复合树脂之间有杂合层形成。结论：UB-Ⅰ粘合剂具有良好的渗透性，与牙本质之间可以形成良好的机械嵌合。