不同性状激光微孔脱细胞真皮构建活性基质的研究

目的：探讨不同性状激光微孔脱细胞真皮构建活性基质的可行性。方法：将激光微孔猪脱细胞真皮分成两组即干燥组和湿润组，每组36片。将成纤维细胞悬液分别滴加在两组真皮表面，培养不同时间点后，每组分别随机留取6个孔的培养液，采用双抗体夹心ELISA法测定上清液中人TGF-β1、bFGF含量。结果：随着培养时间的延长，TGF-p。和bFGF分泌量逐渐增多，第8天达到稳定水平，第10、12天与第8天的含量比较差异无统计学意义（P〉0．05）；各时间点干燥组各因子的含量均高于湿润组（P〈0．05）。结论：激光微孔猪脱细胞真皮可用为支架构建活性基质，其冷冻干燥后更有利于成纤维细胞的粘附与增殖。     

胰岛素治疗对烫伤脓毒症骨骼肌蛋白高降解的影响

目的探讨胰岛素对烫伤脓毒症骨骼肌蛋白高降解的调节。方法沸水烫伤并注射内毒素建立烫伤和脓毒症兔模型,使用胰岛素控制血糖。检测伸趾长肌蛋白降解率和蛋白酶体的基因与蛋白表达变化。结果胰岛素治疗显著降低了伤后骨骼肌蛋白降解率和蛋白酶体的表达。结论胰岛素能抑制泛素-蛋白酶体途径活性的增强,减轻烫伤和脓毒症时骨骼肌蛋白高降解。     

卡巴胆碱对烧创伤后肠道功能障碍影响的研究

目的观察肠道内给予卡巴胆碱对兔肠部分缺血再灌注(I/R)损伤及重度烧伤患者肠道功能障碍的影响。方法将50只大白兔制成肠部分I/R损伤模型后,随机分为肠部分I/R损伤组(25只)、卡巴胆碱组[25只,于肠系膜上动脉(SMA)阻断后1h肠内注入3g/L卡巴胆碱(3μg/kg)]；另取25只设为假手术组,仅分离SMA,不阻断；取5只作为正常对照组,不致伤,处死后留取标本待测。检测兔SMA阻断前后及肠道内给予卡巴胆碱后肠黏膜的血流量。各致伤组均在处理后2、4、6、8、24、48、72h留取静脉血测定其血浆二胺氧化酶(DAO)活性及D-乳酸和D-木糖含量。并行葡聚糖蓝排出实验,以检测胃肠道吸收功能。同时选择大面积烧伤[烧伤总面积(84±12)%TBSA]患者8例,在患者肠呜音＜2次/min或腹胀明显时,口服1g/L卡巴胆碱(15μg/kg),观察给药后每分钟肠鸣音次数及腹胀情况。结果SMA阻断后肠部分I/R损伤组肠黏膜血流量为(48±6)PU,较正常对照组[(102±5)PU]明显减少,而肠道内注入卡巴胆碱后1h血流量增至(77±3)PU。肠缺血后肠部分I/R损伤组血浆DAO活性及D-乳酸含量开始升高,处理后24h达峰值[(4．63±0．27)U/ml、(7．9±2．4)mg/L],以后逐渐下降,但仍高于正常对照组[(0．89±0．14)U/ml、(2．0±1．1)mg/L,P＜0．05]。卡巴胆碱组的变化基本同肠部分I/R损伤组,但变化幅度较小；而假手术组则无明显变化(P＞0．05)。在给予D-木糖后2h,肠部分I/R损伤组血浆D-木糖含量显著降低,但处理后6h肠部分I/R损伤组及卡巴胆碱组明显升高,以后逐渐下降；假手术组略有波动。SMA处理后2h肠部分I/R损伤组葡聚糖蓝未见排出,处理后6h其运动距离逐渐增加,但处理后24h其运动距离仍明显短于正常值(P＜0．05),48～72h基本恢复正常；卡巴胆碱组注入葡聚糖蓝后即可见其排出,其运动距离明显增加,处理后6h达峰值(43±6)cm,以后逐渐缩短接近正常(28±3)cm。给药前患者肠呜音较弱(1．6±1．1)次/min,给药后10 min明显增强为(6．9±1．7)次/min,30 min时为(8．3±2．4)次/min,给药后1h患者肠鸣音仍较活跃,为(6．1±1．3)次/min。给药后2h患者腹胀明显减轻,其中有6例患者开始排便。结论肠内给予卡巴胆碱可增加兔肠黏膜血流量,改善其肠道运动、吸收、屏障功能；大面积烧伤患者口服卡巴胆碱,可改善其肠道功能障碍。     

严重烧伤大鼠早期切痂植皮后骨骼肌泛素转录的变化

目的　探讨大鼠严重烧伤早期切痂后骨骼肌蛋白降解率变化的内在机制。方法　应用大鼠 30 %Ⅲ度背部烫伤模型 ,随机分为伤前、单纯烧伤、早期切痂和非早期切痂组 ,切痂组分别于伤后 1天和 4天行切痂手术 ,采用无创技术和骨骼肌充分氧供离体孵育系统测定伸趾长肌肌纤维蛋白降解率 ,Northernblot方法测定伸趾长肌组织中泛素转录表达的变化。结果　伸趾长肌肌纤维蛋白降解率在伤后各时间点均显著增加 (P<0 0 1) ,早期切痂组较非早期切痂组在伤后 2、4、5天显著降低 (P <0 0 5 ,P <0 0 1)。泛素转录表达在伤后均显著增加 ,但早期切痂组呈持续下降的趋势。蛋白降解率与泛素转录表达间呈显著正相关 (r=0 94 78,P <0 0 5 )。结论　烧伤早期切痂可降低骨骼肌蛋白的高分解代谢反应 ,与泛素 蛋白酶体途径活性下降有关     

腰-硬联合麻醉对剖宫产产妇呼吸参数的影响

目的：探讨腰-硬联合麻醉对剖宫产产妇呼吸功能的影响。方法：30例美国麻醉医师学会（ASA）Ⅰ～Ⅱ单胎足月孕妇,在腰-硬联合麻醉下行剖宫产术,用面罩紧闭法测量孕妇在人室后平卧位（T1）,麻醉完成后仰卧即刻（T1）、5rain（T2）、10min（T3）,胎儿取出后1min（T4）、5min（T5）和10min（T6）时的呼吸频率（RR）、潮气量（VT）、分钟通气量（VE）、呼气末二氧化碳分压（PETCO2）、1秒率（V1．0）、呼吸比（E：I）等参数的变化。结果：与基础值（T0）相比较,PETCO2于各个观察时间点均明显降低（P〈0．01）;T1、T2、T6时VE明显降低;T1时的VT亦降低（P〈0．05）。结论：腰-硬联合麻醉能使剖宫产产妇的VE、PETCO2明显降低,缓解产妇的过度通气,降低其氧耗量,且不会对产妇呼吸功能产生不利影响。     

异体表皮细胞和成纤维细胞库的建立

目的 寻找表皮细胞和成纤维细胞体外培养、鉴定、冻存与复苏的可行方法.方法 酶消化法分离表皮细胞和成纤维细胞,分别进行原代、传代培养,并对细胞进行冻存与复苏.表皮细胞进行广谱角蛋白-异硫氰酸荧光素(PAN-FITC)免疫荧光染色,成纤维细胞行波形蛋白-FITC免疫荧光染色.结果 表皮细胞呈克降样增长,以后细胞逐渐互相融合.PAN-FITC染色均为阳性.成纤维细胞的体外生长活力强,波形蛋白-FITC染色呈阳性.结论 酶消化法是表皮细胞和成纤维细胞体外培养的重要方法,本实验为构建组织工程皮肤奠定了基础.     

卡巴胆碱对烧伤延迟复苏大鼠免疫功能的影响

Objective To explore the changing trend of Ia on monocyte, lymphocyte apoptosis rate, TNF-α and IL-6 in abdominal aorta of burned rats with delayed resuscitation and the influence of application of carbachol on them. Methods Adult male Wistar rats were randomly divided into normal control group(n =8), scald group(n =48) and scald with carbachol treatment group(n =48). In latter two groups, rats were inflicted with 30% total body surface area (TBSA) full-thickness scald and delayed fluid resuscitation. All scald rats were sacrificed at the 6th hours or 1st, 2nd, 3rd, 7th, 14th day after scald, with 8 rats at each time point. Expression of Ia antigen on monocyte and lymphocyte apoptosis rate were determined by direct immunofluorescence on a flow cytometer, and TNF-α and IL-6 was measured by ELISA. Results Expression of la on monocyte was obviously lower than that of controls. The lowest levels were recorded on the 6th hours and 1st day after scald. Subsequently, Ia was elevated gradually, but still lower than that of normal rats(P <0. 01). After administration of carbachol, Ia expression was obviously promoted, compared with the simple scald group (P <0. 01). Lymphocyte apoptosis rate, TNF-α and IL-6 was higher than that of controls(P <0. 01). After administration of cavachol, , lymphocyte apoptosis rate and TNF-α and IL-6 was obviously down-regulated on the 6th hours, 1st day, 2nd day and 3rd day after scald injury, compared with the simple scald group (P < 0. 01 or 0. 05). Conclusion After severe burn with delayed fluid resuscitation, there is a low la expression, high lymphocyte apoptosis rate and increased releasing of proinflammatory cytokine. Immune function was suppressed. Carbacho] could improve the immune function of scald rats with delayed fluid resuscitation.     

卡巴胆碱对烧伤延迟复苏大鼠免疫功能的影响

Objective To explore the changing trend of Ia on monocyte, lymphocyte apoptosis rate, TNF-α and IL-6 in abdominal aorta of burned rats with delayed resuscitation and the influence of application of carbachol on them. Methods Adult male Wistar rats were randomly divided into normal control group(n =8), scald group(n =48) and scald with carbachol treatment group(n =48). In latter two groups, rats were inflicted with 30% total body surface area (TBSA) full-thickness scald and delayed fluid resuscitation. All scald rats were sacrificed at the 6th hours or 1st, 2nd, 3rd, 7th, 14th day after scald, with 8 rats at each time point. Expression of Ia antigen on monocyte and lymphocyte apoptosis rate were determined by direct immunofluorescence on a flow cytometer, and TNF-α and IL-6 was measured by ELISA. Results Expression of la on monocyte was obviously lower than that of controls. The lowest levels were recorded on the 6th hours and 1st day after scald. Subsequently, Ia was elevated gradually, but still lower than that of normal rats(P <0. 01). After administration of carbachol, Ia expression was obviously promoted, compared with the simple scald group (P <0. 01). Lymphocyte apoptosis rate, TNF-α and IL-6 was higher than that of controls(P <0. 01). After administration of cavachol, , lymphocyte apoptosis rate and TNF-α and IL-6 was obviously down-regulated on the 6th hours, 1st day, 2nd day and 3rd day after scald injury, compared with the simple scald group (P < 0. 01 or 0. 05). Conclusion After severe burn with delayed fluid resuscitation, there is a low la expression, high lymphocyte apoptosis rate and increased releasing of proinflammatory cytokine. Immune function was suppressed. Carbacho] could improve the immune function of scald rats with delayed fluid resuscitation.     

大面积侵袭感染组织切除对烧伤创面脓毒症患者静息能量消耗的影响

目的观察大面积侵袭感染组织切除对烧伤创面脓毒症患者高代谢的影响。方法对连续救治的8例烧伤创面脓毒症患者,分别于大面积侵袭感染组织切除前,手术后和病情稳定时,对静息能量消耗(restingenergyexpenditure,REE)白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子α(TNFα)、内毒素(LPS)进行监测。结果侵袭感染组织切除后REE水平[(307.7±31.3)kJ·h     

烫伤大鼠肠道防御素-5和相关酶转录表达及与细菌移位关系的探讨

目的　探讨烧伤对大鼠肠道防御素 5 ( RD 5 )及其活化所需的基质金属蛋白酶 Matrilysin表达的影响及与肠道细菌移位的关系。方法　 SPF级 Wistar大鼠 32只 ,随机分为对照组 (假烫组 ,n=8)和烫伤组 ( n=2 4 )。烫伤组大鼠均在背上造成 30 %总体表面积 度烫伤。伤后 8、2 4和 72 h切取末端回肠 ,利用逆转录聚合酶链反应 ( RT PCR)方法测定不同时间点 RD 5 m RNA及 Matrilysin m RNA表达变化。伤后 2 4 h同时取肠系膜淋巴结 ( ML N )、肝、脾、肺组织进行细菌培养 ,计算脏器细菌移位频率 ;取回肠观察帕内特细胞形态。结果　 RD 5 m RNA表达量在伤后 8h显著升高 ( P<0 .0 5 ) ,伤后 2 4 h降低 ,但仍高于对照组水平 ,而伤后 72 h则降至对照组以下 ;Matrilysin m RNA在伤后 8h和 2 4 h略有增高 ,但伤后 72 h表达显著增强( P<0 .0 5 )。伤后 2 4 h脏器细菌移位频率 ( 5 8.3% )显著高于对照组 ( 8.3% ) ,P<0 .0 1;帕内特细胞无明显形态学改变。结论　大鼠烫伤后早期肠道 RD 5 m RNA及 Matrilysin m RNA表达增强 ,但时相略有不同。这可能是机体对抗细菌移位的一种保护性反应