大鼠急性β射线皮肤损伤创面愈合过程中Ang-1及其受体的表达

目的探讨急性β射线皮肤损伤创面难愈合过程中血管生成素1(Ang-1)及其受体Tie-2表达的动态变化。方法雌性大鼠54只分为3组:照射组(n=24),直线加速器产生β射线(45Gy)单次照射大鼠臀部皮肤40mm×40mm,建立急性β射线皮肤损伤模型;烧伤组(n=24),以直径为25mm的恒温电烙铁,温度80℃,时间8s,建立大鼠深Ⅱ度热力烫伤动物模型;对照组(n=6),正常大鼠。取不同时期创面皮肤组织,采用免疫组化SP法及RT-PCR等方法,检测1、2、3、4周不同时相点各组大鼠创面组织中Ang-1蛋白、Tie-2蛋白和Ang-1mRNA、Tie-2mRNA的表达。结果正常大鼠皮肤中Ang-1、Tie-2积分吸光度(A)值低表达;照射组伤后1、2和3周的Ang-1 A值分别为0.130±0.012、0.170±0.013和0.192±0.072,明显低于烧伤组的0.190±0.182、0.332±0.014和0.273±0.014(P<0.05);照射组1、2和3周的Tie-2A值分别为0.112±0.011、0.183±0.012和0.171±0.061,亦明显低于烧伤组的0.170±0.062、0.310±0.013和0.284±0.014(P<0.05);Ang-1mRNA、Tie-2mRNA表达的变化与Ang-1蛋白、Tie-2蛋白相似。结论急性β射线皮肤损伤创面愈合过程中,Ang-1及Tie-2持续低表达可能是急性β射线皮肤损伤创面难愈合的机制之一。     

大鼠急性β射线皮肤损伤创面愈合过程中PDGF及其受体的表达

目的 研究血小板源性生长因子A(PDGF-A)及其受体(PDGFR-α)在大鼠急性β射线皮肤损伤创面愈合过程中的变化.方法 雌性SD大鼠54只,随机分为3组:照射组(n=24),直线加速器产生的β射线(45Gy)单次照射大鼠臀部皮肤20mm×40mm,建立急性深Ⅱ°β射线皮肤损伤动物模型;烫伤组(n=24),以直径为25 mm的恒温电烙铁,温度80℃,时间8 s,建立大鼠深Ⅱ°热力烫伤动物模型;对照组(n=6),正常大鼠.采用免疫组织化学观察创面出现后0、1、2、3、4、8周符组大鼠创面组织中PDGF-A/PDGFR-α表达.结果 照射组大鼠创面出现后1、2、3周创面组织中PDGF-A和PDGFR-α表达均为阳性,0、4、8周表达为弱阳性.烫伤组烫伤后2周创面组织中PDGF-A和PDGFR-α表达均为强阳性,0、1、3周表达为阳性,4、8周表达为弱阳性.对照组大鼠正常皮肤中PDGF-A及PDGFR-α表达为弱阳性.结论 急性β射线皮肤损伤创面愈合过程中PDGF-A及其受体PDGFR-α表达减弱,并持续低表达,不能形成峰值,可能是急性β射线皮肤损伤创面难愈合的机制之一.     

大鼠电子射线皮肤损伤动物模型的建立

目的采用电子射线每周重复照射建立大鼠放射性皮肤损伤模型,探讨其科学性、可行性及实用性。方法 36只Wistar雌性大鼠,按5∶1比例随机分为照射组(A组)30只、对照组(B组)6只。采用6MV电子线对A组大鼠左侧臀部皮肤进行分次照射(5Gy/次,1次/周,累积剂量最高为60Gy),于照射后第1、4、8、12、16周处死,每时相点处死6只,B组于第2周末处死。观察动物活体外观、皮肤的大体标本、行苏木素-伊红染色观察大鼠皮肤组织学变化。结果 A组受照皮肤组织表现出放射性皮炎及皮肤纤维化的变化过程。结论每周大剂量电子射线重复照射所制作的大鼠放射性皮肤损伤模型稳定可靠,符合临床放疗实际。     

rhGM-CSF和纳米银联合外用对深Ⅱ°烫伤创面上皮化的影响

目的研究重组人粒细胞-巨噬细胞集落刺激因子(rh GM-CSF)和纳米银敷料联合外用对深Ⅱ°烫伤创面愈合过程上皮化的影响。方法用Wistar大鼠建立深Ⅱ°烫伤模型120例,均分为4组,A组用凡士林纱布覆盖、B组用纳米银敷料覆盖、C组用rh GM-CSF涂抹创面、D组用rh GM-CSF+纳米银敷料覆盖,伤后第1、4、7、10、14、21天,观察创面的病理学改变,用ELISA法测定血清中的表皮生长因子(EGF),采用RT-PCR法检测创面愈合过程中TGF-β1mRNA表达的变化。结果 4个组均在第10天出现明显的上皮化;EGF水平逐渐上升,A组上升最慢,D组最快;A、B组的TGF-β1mRNA相对表达量逐渐上升,C、D组的在第14天达到峰值,第21天下降,且D组下降幅度较大。结论 rh GM-CSF和纳米银联合外用,加速了深Ⅱ°烫伤创面愈合过程中上皮化的形成,上皮化程度优于单独应用。     

重组人粒细胞-巨噬细胞集落刺激因子对深Ⅱ度烫伤模型大鼠创面愈合的影响

目的研究外用重组人粒细胞-巨噬细胞集落刺激因子(rh GM-CSF)对深Ⅱ度烫伤创面愈合的影响。方法90只Wistar大鼠建立深Ⅱ度烫伤模型,分为A、B、C三组,每组30只。A组:凡士林纱布覆盖;B组:纳米银敷料覆盖;C组:rh GM-CSF涂抹创面。伤后第1,4,7,10,14,21天,观察创面病理学改变,按照酶联免疫吸附法测定血清中表皮生长因子(EGF)水平,运用逆转录-聚合酶链反应(RT-PCR)检测创面愈合过程中转化生长因子(TGF)-β1mRNA表达的变化。结果病理学改变:A、B、C三组均在第10天出现明显的上皮化;EGF水平逐渐上升,A组上升最慢,C组上升最快,第1,4天各组间及第7天A组与B组差异无统计学意义(P>0.05),第7天其余各组间及第10,14,21天各组间差异均有统计学意义(均P<0.05);TGF-β1mRNA相对表达量,A组、B组逐渐上升,C组在第14天达到峰值,第21天下降,第1天,B组与C组之间差异有统计学意义(P<0.05),第4天各组间差异无统计学意义(P>0.05),第7,10,14,21天各组间差异有统计学意义(P<0.05)。结论 rh GM-CSF可加速深Ⅱ度烫伤创面愈合。     

复方竹叶石膏颗粒对家兔急性放射性食管损伤Bcl-2与Bax mRNA及蛋白表达的影响

目的 探讨复方竹叶石膏颗粒对家兔急性放射性食管炎组织中Bax和Bcl-2 mRNA及蛋白表达的影响.方法 将40只雄性家兔随机分为空白对照组(A组)、单纯照射组(B组)、复方竹叶石膏颗粒组(C组)、康复新液组(D组),每组10只.B、C、D组均用6 mV-X射线对家兔食管单次照射32 Gy.C、D组于照射前1周照射至放疗结束后7d.4组均于照射后第7天留取家兔全长食管,检测Bax和Bcl-2 mRNA和蛋白的表达水平.结果 B组家兔食管组织中Bax mRNA与蛋白表达水平较A组升高,Bcl-2 mRNA与蛋白表达及Bcl-2/Bax值较A组降低(P＜0.05).C、D组Bcl-2 mRNA与蛋白表达水平及Bcl-2/Bax值高于B组,Bax mRNA与蛋白表达水平低于B组(P＜0.05).C组Bcl-2 mRNA与蛋白表达水平均高于D组(P＜0.05).结论 复方竹叶石膏颗粒能够防治急性放射性食管炎,其机制可能与上调Bcl-2基因与下调Bax基因的表达有关.     

伤疡愈软膏对烫伤大鼠创面碱性成纤维细胞生长因子表达水平的影响

目的:探讨伤疡愈软膏对烫伤大鼠创面愈合过程中碱性成纤维细胞生长因子(b-FGF)表达水平的影响。方法:选择SD大鼠60只,随机分为A、B、C组,每组20只。A组为正常对照组,B、C组复制烫伤动物模型,其中B组大鼠经常规处理,C组大鼠使用伤疡愈软膏治疗。愈合过程中取组织溶浆液,应用逆转录聚合酶反应检测烫伤创面b-FGF mRNA的表达水平。结果:与A组比较,B、C组大鼠创面b-FGF mRNA的表达水平有显著差异;与B组比较,C组大鼠烫伤创面愈合时间显著缩短。结论:伤疡愈软膏可促进创面愈合,提高b-FGF mRNA的表达水平。     

益气活血中药对放射性肺损伤大鼠NOS3、MTHFR表达的影响

目的：探讨益气活血方防治放射性肺损伤的分子生物学机制和相关敏感因子。方法将108只雌性SD大鼠采用随机数字表法分为中药组(A组)、单纯照射组(B组)及健康对照组(C组),每组36只。应用6MV-X直线加速器对前2组大鼠右肺进行照射(5 Gy/次,2次/周,累积剂量为30 Gy),A组于受照射当天开始用益气活血方1 ml灌胃、2/d,B组及C组用蒸馏水1 ml灌胃、2/d,停止时间为照射结束后2周。各组于照射开始第2、4、8、12、16、20周末随机抽取6只大鼠处死,取其肺组织,用反转录-聚合酶链反应( RT-PCR)及免疫组化方法检测大鼠肺组织内皮型一氧化氮合酶(NOS3)、亚甲基四氢叶酸还原酶(MTHFR)mRNA及蛋白表达。结果 A组与B组NOS3、MTHFR的mRNA及蛋白表达于照射后第2周开始增高,第8周达高峰,第12周开始下降,16周后降低明显,与C组相比,差异均有统计学意义(P<0．01);A组与B组相比,NOS3、MTHFR mRNA及蛋白表达趋势类似,A组表达水平在12周之前均高于B组,差异有统计学意义(P<0．05,P<0．01)。结论 NOS3、MTHFR可能为放射性肺损伤的敏感因子。益气活血方可通过上调NOS3、MTHFR的表达起到减轻放射性肺损伤的作用。临床放射性肺损伤治疗时间应延长2~3个月,各时间段治疗重点应有所侧重。     

神经肽P物质对放射损伤皮肤成纤维细胞分泌功能及VEGF、bFGF、PDGF表达的影响

目的 探讨神经肽P物质（SP）对放射损伤皮肤成纤维细胞分泌功能及血管内皮生长因子（VEGF）、碱性成纤维细胞生长因子（bFGF）、血小板衍生生长因子（PDGF）表达的影响。方法 将对数生长期皮肤成纤维细胞HFF-1按照射剂量分为未照射组（0 Gy）,照射组（2 Gy、6 Gy、12 Gy、18 Gy）,SP干预组（0 Gy+SP、2 Gy+SP、6 Gy+SP、12 Gy+SP、18 Gy+SP）。照射后24 h,酶联免疫吸附试验检测培养基及细胞裂解液Ⅰ型胶原（Col-Ⅰ）、Ⅲ型胶原（Col-Ⅲ）、降钙素基因相关肽（CGRP）、VEGF、bFGF、PDGF表达情况;实时荧光定量PCR检测细胞VEGF、bFGF、PDGF mRNA表达。结果 与0 Gy组比较,培养基Col-Ⅰ、Col-Ⅰ/Col-Ⅲ及细胞内CGRP、VEGF蛋白在各照射组均升高;bFGF、PDGF mRNA在照射组（6 Gy、12 Gy、18 Gy）升高;Col-Ⅰ在6 Gy组培养基与细胞内一致高表达;Col-Ⅰ、Col-Ⅲ、Col-Ⅰ/Col-Ⅲ、CGRP在18 Gy组培养基与细胞内均一致高表达。bFGF在照射组（12 Gy、18 Gy）呈现蛋白和mRNA水平高表达。照射前给予SP干预,与未干预相应组相比,CGRP在0 Gy+SP组培养基与细胞内一致高表达;Col-Ⅰ和Col-Ⅰ/Col-Ⅲ分别在干预组（2 Gy+SP、6 Gy+SP）和（2 Gy+SP、6 Gy+SP、18 Gy+SP）培养基与细胞内一致低表达;VEGF和PDGF分别在干预组（2 Gy+SP、12 Gy+SP、18 Gy+SP）和（12 Gy+SP、18 Gy+SP）呈现蛋白和mRNA水平一致高表达。结论 SP可调节放射损伤皮肤成纤维细胞Col-Ⅰ、Col-Ⅲ表达与分泌,降低Col-Ⅰ/Col-Ⅲ比值,协同成纤维细胞对电子束照射的应激反应,促进放射损伤皮肤成纤维细胞中VEGF、PDGF的高表达。     

益气活血中药对大鼠放射性肺损伤TGF-β1蛋白表达的调控作用

目的:观察益气活血中药对小剂量重复照射下不同时相大鼠血清、肺组织中转化生长因子-β1(TGF-β1)蛋白表达的影响,探讨其对放射性肺损伤的防治作用.方法:将96只Wistar雌性大鼠随机分为照射加中药组(A组)30只、照射加氨溴索组(B组)30只、单纯照射组(C组)30只、正常对照组(D组)6只.用6-Mv直线加速器对前3组大鼠右肺进行分次照射(每次5 Gy,1次/周,累积剂量最高为30 Gy),A组每日予自制益气活血中药1 mL灌胃,B组每日子氨溴索(50 mg/kg)灌胃,于照射后第4、6、8、12、26周末分别处死每组各6只大鼠,D组6只于第4周末处死,取血清标本进行ELISA检测.取肺组织标本进行免疫组化检测,观察各组不同时相血清、肺组织中TGF-β1蛋白表达的动态变化.结果:3组血清、肺组织中TGF-β1蛋白表达于照射后第4周升高,第12周达高峰,第26周略下降;A组与B组各时相TGF-β1蛋白表达水平近似(P>0.05),但均低于C组(P<0.05).结论:益气活血中药具有抑制TGF-β1合成分泌的作用,可能是其具有减轻早期放射性炎症反应和后期放射性肺纤维化作用的机制.     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg.kg) or i.p. (50 mg.kg) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) l.h. kg in the male rat and 10.6 (95% CI: 7.5, 15.0) l.h. kg in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p 0.001) in plasma obtained from the male (8.8 2.0%) compared with the female rat (11.7 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

休克时血中氧自由基的变化

本实验测定10名休克患者血浆和红细胞的丙二醛(MDA)、血浆总抗的氧化活性(AOA)的含量。结果表明:休克病人红细胞膜和血浆 MDA 含量(4.298±0.722;5.348±0.834)与对照组(3.235±0.682;4.356±1.081)比较明显增高(P<0.05);血浆 AOA(39.65±7.858)与对照组(48.21±10.81)比较明显降低(P<0.01)。提示:休克时,患者机体内自由基反应增强是引起组织细胞损伤的原因之一。     

Quantitative in vivo and in vitro sex differences in artemisinin metabolism in rat

1. The pharmacokinetics of the antimalarial compound artemisinin were compared in the male and female Sprague-Dawley rat after single dose i.v. (20 mg x kg(-1)) or i.p. (50 mg x kg(-1)) administration of an emulsion formulation. 2. Plasma clearance of artemisinin was 12.0 (95% confidence interval: 10.4, 13.0) 1 x h(-1) x kg(-1) in the male rat and 10.6 (95% CI: 7.5, 15.0) 1 x h(-1) x kg(-1) in the female rat suggesting high hepatic extraction in combination with erythrocyte uptake or clearance. Artemisinin half-life was approximately 0.5 h after both routes of administration in both sexes. Values for plasma clearance and half-lives did not statistically differ between the sexes. 3. After i.p. administration artemisinin AUCs were 2-fold higher in the female compared with male rat (p < 0.001). Artemisinin disappearance was 3.9-fold greater in microsomes from male compared with female livers and it was inhibited in male microsomes by goat or rabbit serum containing antibodies against CYP2C11 and CYP3A2 but not CYP2B1 or CYP2E1. 4. The unbound fraction of artemisinin in plasma was lower (p < 0.001) in plasma obtained from the male (8.8 +/- 2.0%) compared with the female rat (11.7 +/- 2.2%). 5. The possibility of a marked sex difference, dependent on the route of administration, has to be taken into account in the design and interpretation of toxicological studies of artemisinin in this species.     

Theophylline kinetics in peripheral tissues in vivo in humans

Several biochemical and cellular effects have been described for methylxanthines under in vitro conditions. However, it is unknown, whether threshold concentrations required to exert these effects are attained in target tissues in vivo. We therefore employed the microdialysis technique for measuring theophylline concentrations in peripheral tissues under in vivo conditions.Following in vitro and in vivo calibration, microdialysis probes were inserted into the medial vastus muscle and into the periumbilical subcutaneous adipose layer of healthy volunteers. Following single oral dose administration of 300 mg or i.v. infusion of 240 mg theophylline, in vivo time courses of theophylline concentrations were monitored in tissues and plasma. Major pharmacokinetic parameters (c_max, t_max, AUC) were calculated for plasma and tissue time courses. The mean AUC_tissue /AUC_plasma-ratio was 0.56 (p.o.) and 0.55 (i.v.) for muscle and 0.55 (p.o.) and 0.72 (i.v.) for subcutaneous adipose tissue.We conclude that microdialysis provides important information on the distribution and the tissue pharmacokinetics of theophylline.Abbreviations FPIA Fluorescence polarisation immuno assay - AUC Area under the curve - t_max Time to peak concentration - c_max Peak concentration     

Interindividual variability in metabolic activation in humans in vivo

In assessing interindividual variability in metabolic activation, the toxic metabolite is often too unstable for conventional analysis. Possible alternatives include a stable product of the reactive metabolite e.g. cysteinyl derivatives of N-acetyl-4-benzoquinoneimine, the toxic metabolite of paracetamol, adducts with DNA or protein, and indirect measurement of the activity of the enzyme(s) producing the active metabolite. An example of the last approach is the use of furafylline, a highly specific inhibitor of human CYP1A2, to determine the extent of the metabolic activation of the cooked food mutagens PhIP and MeIQx. The extent of inhibition, determined from levels of unchanged amine in urine, is an indirect measure of the activity of the activation pathway. Further refinement of this approach, allied to improved measures of the biological process of interest should prove of value in evaluating interindividual variability and its role in the risk assessment process.