烧伤、冲击伤及烧冲复合伤后大鼠早期血清中性粒细胞弹性蛋白酶的变化及意义

Objective To explore the early changes in serum neutrophil elastase(NE) in rats with burn.blast injury or combined bum-blast injury and its significance.Methors A total of 176 male Spragne Dawley rats were randomly divided into four groups:control(C),burn(BU),blast injury(BL) and burn-blast combined injury(BB).Rats in C group were not injured.Animals in BU group were subjected to 25% TBSA full-thickness burn on back with 94℃ water for 12 seconds;Animals in BL group were inflicted with moderate blast injury with 5g 8701 compressed dynamite stick as the explosion source 75 cm away while left chest facing the explosive source;Rats in BB group were burned immediately after the blot injury similarly as in BL group.During the first 24 h post-injury,animals in BU and BB groups received intraperitoneal bronchoalveolar lavage fluid(BALF),water content of lung tissue and NE content in serum were determined at 0 h(C group),3 h,6 h,12 h,1 d,2 d,3 d,7 d post-injury.Results Protein concentration in BALF,water content of lung tissue and NE content in serum in SD rats of the injured groups were significantly higher than those in C group(P＜0.01 or P＜0.05),peaked within 2 d post-injury,especially at 2 d post-injury(NE content in serum:BU group,319.85±19.50,ng/ml;BL group,467.43±31.64 ng/ml;BB group,626.00±26.38 ng/ml vs.C group,78.53±25.10 ng/ml).Overall,protein concentration in BALF.water content of lung tissue and NE content in serum in BB group were significantly higher than BU and BL groups(P＜0.01 or P＜0.05).Correlation analysis showed that within 3 d post-injury.a significant positive correlation was found between the protein concentration in BALF,water content of lung tissue and NE content in serum(r=0.7910,0.8078,P＜0.05) in BU group.NE content in serum and protein concentration in BALF were significantly positively correlated in BB group(r=0.8672,P＜0.05).Conclusion NE may play an important role in early lung injury of burn or blast injury,especially in combined bum-blast injury.     

人血白蛋白的功能及其在危重病治疗中的应用

人血白蛋白(HSA)是一种小分子量、非糖基化、带负电荷的血清蛋白,具有物质结合转运、协调维持血管内皮完整、抗氧化、抗炎、脏器功能保护、维持胶体渗透压等多种功能.综合分析现有的研究及Meta分析结果,HSA用于危重症治疗,可降低并发症发生率,对某些疾病如脓毒症、肝硬化合并自发性腹膜炎等,可显著改善患者预后,但不能确定其可降低危重病患者的病死率.     

高原地区后勤部队军人健康状况的调查

目的调查某高原地区后勤部队军人的健康状况,为制定改善其健康状况的措施提供依据及建议。方法应用自测健康评定量表(self-rated Health measurement scale,SRHMS),采取随机整群抽样法对2011年11月驻扎青海某高原地区的后勤部队共348人进行问卷调查。结果除正向情绪、认知功能、社会健康总分及其社会支持分项外,高原军人SRHMS量表各维度、子量表的均分和总均分均低于一般人群(均P<0.01);在生理健康方面,不同性别、年龄、婚姻状况、兵龄、入伍前居住地的得分差异均有统计学意义(P<0.05或P<0.01);在心理健康方面,不同部别、年龄、婚姻状况、兵龄的得分差异均有统计学意义(均P<0.05);在社会健康方面,不同性别、兵龄的得分差异有统计学意义(P<0.05或P<0.01)。生理健康得分与兵龄有相关性(P<0.05),社会健康得分与性别、文化、部别有相关性(均P<0.05),其余各项间均无相关性(均P>0.05)。结论高原地区后勤部队军人健康状况处于中上水平,但多数项目仍低于一般人群;男性军人、已婚军人、年龄≥30岁的军人、兵龄≥10年的军人应视为重点保健对象;建议高原军队领导应重视健康管理和建立规范的军人健康自测系统,加强有针对性的心理健康教育和健康指导。     

人CD112(Nectin2/PRR2)单克隆抗体的制备和鉴定

目的：表达和纯化CD112 胞膜外区重组蛋白, 制备针对CD112胞膜外区的单克隆抗体（mAb）, 并且研究CD112的分布.方法：采用基因重组技术在真核系统表达CD112-Fc融合蛋白, 亲和层析法进行蛋白纯化;纯化的CD112-Fc融合蛋白免疫BALB/c小鼠, 杂交瘤技术建立分泌CD112 mAb的杂交瘤细胞株, 间接ELISA、 Western blot及FCM鉴定mAb 的特异性.用流式细胞术（FCM）检测 CD112 的细胞系分布.结果：构建了高效真核表达载体pIg3C-CD112, 表达并纯化了CD112-Fc融合蛋白, 其纯度大于 90%.以纯化重组蛋白为免疫原共制备9株分泌抗CD112 mAb的杂交瘤细胞株（命名为FMU-CD112.1 ～FMU-CD112.9）, 制备腹水并纯化 mAb.FMU-CD112.1、 3、 6和8能用于Western blot, FMU-CD112.1、 4、 6和8可用于 FCM.CD112细胞分布检测显示该分子主要分布在正常上皮及内皮细胞系、巨核细胞谱系和部分T细胞系, 并高表达于上皮及内皮来源的肿瘤、胶质瘤等细胞系.结论：成功地表达纯化了CD112-Fc融合蛋白并建立了稳定分泌CD112 mAb的杂交瘤细胞株, 为CD112的功能研究打下坚实的基础.     

人CD226分子胞膜外区D1和D2真核表达载体的构建、表达和鉴定

目的 构建含有人CD226分子胞膜外区结构域1(D1)和结构域2(D2)的真核表达载体,表达并纯化重组蛋白.方法用特异性引物分别扩增CD226 D1和D2的基因,定向插入真核表达载体pSecTag2B-Fc,进行酶切和DNA测序鉴定,重组质粒瞬时转染293T细胞,收集上清,纯化蛋白及SDS-PAGE和Western blot鉴定表达产物.结果利用PCR方法克隆出CD226胞膜外区D1和D2的基因,并正确插入pSecTag2B-Fc载体中,真核表达载体瞬时转染293T细胞,Western blot结果证实转染293T细胞的培养上清液中含有hCD226D1-Fc和hCD226D2-Fc融合蛋白,培养上清经亲和层析柱纯化,可获得较高纯度的重组蛋白.结论 成功的构建了分泌型融合蛋白hCD226D1-Fc和hCD226D2-Fc真核表达载体,获得纯度较高的融合蛋白,为进一步探讨其配受体的相互作用机制奠定了基础.     

三维扫描技术精确控制大鼠烫伤面积动物模型的建立

目的 建立一种可精确控制烫伤面积的大鼠模型.方法 40只大鼠被随机分为5组,每组8只,根据接触94℃热水时间不同分别为6 s组、8 s组、10 s组、12 s组、14 s组,以热水接触作为致伤原因,持续时间控制烫伤深度.比较各组创面烫伤深度和愈合时间,并经病理证实.另取9只大鼠作为3D扫描组,接触94℃热水12 s,烫伤24 h后应用3D流动式双目三维扫描技术,扫描计算大鼠体表面积及烫伤部位占体表总面积的百分比,并与实体解剖测量结果进行比较.结果 12 s组烫伤创面深度可达（34.2±2.9）mm,且无毛囊及汗腺附件存在.创面愈合时间为（24.4±2.2）d,病理证实烫伤深度为深Ⅱ度～Ⅲ度;流动式双目三维扫描技术确认的烫伤面积与实体解剖测量所得结果差异无统计学意义（P〉0.05）.结论 以热水接触为致伤原因,配合三维扫描技术测量烫伤面积精确可信,该模型可以控制烫伤深度、精确烫伤面积,是研究烧（创）伤修复机制及评价创面用药的良好动物模型.     

腺苷一磷酸活化蛋白激酶在严重烫伤大鼠骨骼肌萎缩中的作用

目的:探讨严重烫伤大鼠骨骼肌中腺苷一磷酸活化蛋白激酶（AMPK）的表达和磷酸化水平变化及其在严重烫伤大鼠骨骼肌萎缩中的作用。方法:采用实验研究方法。将100只6周龄雄性Wistar大鼠按随机数字表法分为假伤组和烫伤组,每组50只。测量2组大鼠体重后,烫伤组背部造成30%体表总面积Ⅲ度烫伤,假伤组模拟致烫伤。伤后6 h及...     

应用稳定同位素技术研究烧伤大鼠蛋白质代谢变化

目的 本研究旨在应用稳定同位素技术,在建立稳定的严重烫伤大鼠模型的基础上,量化严重烫伤大鼠伤后总体蛋白代谢率.方法 采用严重烫伤动物模型,雄性SD大鼠20只,随机分为两组:假伤组(n=10)和烫伤组(n=10),伤后4 d给予示踪氨基酸(2H标记的亮氨酸),给予示踪氨基酸后0.5 h,由腹腔动脉取血液标本.将血清样本沉淀取蛋白,分别进行亮氨酸衍生、酮异己酸(KIC)衍生、结合氨基酸处理,经过GC-MC检测,计算比较两组大鼠血浆游离氨基酸2H丰度、亮氨酸更新速率、整体蛋白质分解速率、整体蛋白合成速率.结果 (1)烫伤组血浆游离氨基酸2H丰度明显高于假伤组,烫伤组较假伤组增加了48.24%,差异有统计学意义(P<0.05).(2)烫伤组亮氨酸更新速率为(236.2±13.1)μmol·kg-1·h-1,假伤组为(132.2±6.5)μmol·kg-1·h-1,烫伤组较假伤组增加了78.6%,差异有统计学意义(P<0.05).(3)烫伤组大鼠整体蛋白分解速率为(9.1±1.6)g·kg-1·d-1,较假伤组(5.1±0.5)g·kg-1·d-1有明显增加,前者较后者增高78.4%(P<0.05).(4)烫伤组大鼠整体蛋白合成速率为(5.1±1.8)g·kg-1·d-1,假伤组(4.8±1.1)g·kg-1·d-1,差异无统计学意义(P>0.05).(5)烫伤大鼠为负氮平衡状态[(-4.0±1.0)g·kg-1·d-1].结论 严重烫伤会导致大鼠蛋白质分解速率增加,处于负氮平衡状态.     

烧伤、冲击伤及烧冲复合伤后大鼠早期血清中性粒细胞弹性蛋白酶的变化及意义

Objective To explore the early changes in serum neutrophil elastase(NE) in rats with burn.blast injury or combined bum-blast injury and its significance.Methors A total of 176 male Spragne Dawley rats were randomly divided into four groups:control(C),burn(BU),blast injury(BL) and burn-blast combined injury(BB).Rats in C group were not injured.Animals in BU group were subjected to 25% TBSA full-thickness burn on back with 94℃ water for 12 seconds;Animals in BL group were inflicted with moderate blast injury with 5g 8701 compressed dynamite stick as the explosion source 75 cm away while left chest facing the explosive source;Rats in BB group were burned immediately after the blot injury similarly as in BL group.During the first 24 h post-injury,animals in BU and BB groups received intraperitoneal bronchoalveolar lavage fluid(BALF),water content of lung tissue and NE content in serum were determined at 0 h(C group),3 h,6 h,12 h,1 d,2 d,3 d,7 d post-injury.Results Protein concentration in BALF,water content of lung tissue and NE content in serum in SD rats of the injured groups were significantly higher than those in C group(P＜0.01 or P＜0.05),peaked within 2 d post-injury,especially at 2 d post-injury(NE content in serum:BU group,319.85±19.50,ng/ml;BL group,467.43±31.64 ng/ml;BB group,626.00±26.38 ng/ml vs.C group,78.53±25.10 ng/ml).Overall,protein concentration in BALF.water content of lung tissue and NE content in serum in BB group were significantly higher than BU and BL groups(P＜0.01 or P＜0.05).Correlation analysis showed that within 3 d post-injury.a significant positive correlation was found between the protein concentration in BALF,water content of lung tissue and NE content in serum(r=0.7910,0.8078,P＜0.05) in BU group.NE content in serum and protein concentration in BALF were significantly positively correlated in BB group(r=0.8672,P＜0.05).Conclusion NE may play an important role in early lung injury of burn or blast injury,especially in combined bum-blast injury.     

人脐带间充质干细胞促进微粒皮复合异体皮移植修复兔皮肤创面的实验研究

目的观察人脐带间充质干细胞（hUC-MSC）与大耳兔自体微粒皮复合同种异体皮移植促进修复其皮肤缺损创面的效果。方法取成年日本大耳兔8只,构建兔皮肤缺损模型,每2只兔交换移植反削断层皮,异体真皮面均匀黏附兔自体微粒皮。头侧创面为hUC-MSC移植组（A组）、尾侧创面为PBS液空白对照组（B组）。术后21 d观察创面情况,计算创面愈合率,进行统计学分析;术后28 d切取创面愈合区域组织,对标本行苏木精-伊红染色观察。结果术后21 d创面愈合率A组（85.1±4.0）%,高于B组（79.5±4.3）%,两组比较差异有统计学意义（P〈0.05）。取材病理学观察可见两组组织均缺少皮肤附件;A组表皮层可发现表皮钉突样结构,而B组表皮层底部平坦,与基底结合较为疏松。结论推测hUC-MSC可促进创面微粒皮生长、扩展,缩短创面愈合时间,提高愈合质量。