半乳糖凝集素-1预处理对机械通气相关性肺损伤小鼠的影响

目的 探讨半乳糖凝集素-1(Galectin-1)预处理对机械通气相关性肺损伤(VILI)小鼠的影响。方法 选择清洁级健康雄性C57BL/6小鼠30只,6～8周龄,体重22～30 g。采用随机数字表法将小鼠分为三组：对照组(C组)、VILI组(V组)和Galectin-1+VILI组(G组),每组10只。C组气管插管后保持自主呼吸4 h, V组和G组气管插管后机械通气4 h。气管插管前1 h C组和V组腹腔注射生理盐水0.75 ml, G组腹腔注射Galectin-1 3μg。于气管插管前即刻、自主呼吸或机械通气结束时采集动脉血检测PaO₂,后处死小鼠,收集肺泡支气管灌洗液(BALF),采用ELISA法检测BALF中IL-1β和IL-18浓度。取肺组织测定湿/干重比(W/D),采用qRT-PCR法检测肺组织GSDMD、caspase-1和caspase-11 mRNA表达量,Western blot法检测肺组织GSDMD、caspase-1和caspase-11蛋白含量,HE染色法观察病理改变并行肺损伤评分。结果 与C组比较,V组和G组机械通气结束时PaO     

组织蛋白酶B在大鼠呼吸机相关性肺损伤中的作用：与NLRP3炎症小体的关系

目的评价组织蛋白酶B(CTSB)在大鼠呼吸机相关性肺损伤(VILI)中的作用及其与NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体的关系。方法 SPF级健康雄性SD大鼠36只, 6～8周龄, 体重220～300 g, 采用随机数字表法分为3组(n=12):对照组(C组)、VILI组(V组)和VILI+CA074-me组(Me组)。C组和V组大鼠气管插管前1 h腹腔注射等量生理盐水, Me组腹腔注射CA074-me 5 mg/kg。C组自主呼吸4 h, V组和Me组行机械通气4 h, 通气参数:VT 20 ml/kg, 通气频率80次/min, FiO2 21%, PEEP 0 cmH2O。气管插管前和自主呼吸或通气结束后采集股动脉血行动脉血气分析, 记录PaO2, 随后处死大鼠, 收集支气管肺泡灌洗液(BALF)和取肺组织, 确定肺组织湿重/干重(W/D)比值, HE染色法观察病理学结果并进行肺损伤评分, 采用ELISA法检测BALF及血清IL-1β和IL-18浓度。采用qRT-PCR法检测肺组织CTSB、NLRP3、凋亡相关斑点样蛋白(ASC)和caspase-1的mRNA表...     

吸入七氟醚、地氟醚对大鼠肺组织超微结构和肺表面活性物质的影响

目的观察不同吸入浓度七氟醚和地氟醚对大鼠肺组织超微结构及支气管肺泡灌洗液 (BALF)中肺表面活性物质相关蛋白-A(SP-A)、磷脂酰胆碱(PC)浓度的影响。方法雄性Wistar大鼠 50只,随机分为5组,每组10只,C组:单纯机械呼吸组;S1组和S2组:分别吸入1．0 MAC、1．5 MAC七氟醚4 b;D1组和D2组:分别吸入1．0MAC、1．5MAC地氟醚4 b。在相应时点处死大鼠,观察肺组织超微结构并测定BALF中SP-A和PC浓度。结果与C组比较,S1、S2、D1、D2组肺泡Ⅱ型上皮细胞微绒毛破坏程度较重,以D1、D2组改变更明显,板层体减少,并呈大量空泡化;BALF中SP-A、PC浓度下降 (P<0．01或0．05)。结论吸入1-0 MAC、1．5 MAC七氟醚或地氟醚4 h可降低肺泡Ⅱ型上皮细胞合成肺表面活性物质。     

幼猪机械通气中异氟醚或七氟醚混合一氧化氮吸入的安全性

目的研究吸入异氟醚或七氟醚混合一氧化氮(NO)在幼猪机械通气中的安全性。方法36头幼猪随机分为6组:Ⅰ组(对照组):单纯机械通气;Ⅱ组(NO组):吸入20ppmNO;Ⅲ组(异氟醚组):吸入1.3MAC异氟醚;Ⅳ组(异氟醚 NO组):吸入1.3MAC异氟醚及20ppmNO;Ⅴ组(七氟醚组)吸入1.3MAC七氟醚;Ⅵ组(七氟醚 NO组)吸入1.3MAC七氟醚及20ppmNO。用麻醉机行间歇正压通气4h,测定各组机械通气前、机械通气1、2、3、4h(T0、T1、T2、T3、T4)的呼吸频率(RR)、呼吸系统总顺应性(Crs)、气道压力(Paw)、潮气量(VT)、分钟通气量(MV)以及呼末二氧化碳分压(PETCO2);测定T0、T2、T4时点动脉血高铁血红蛋白(MetHb)和亚硝酸根(NO2-/NO3-)水平;处死动物后比较各组肺组织湿/干重比、支气管肺泡灌洗液(BALF)中饱和磷脂/总磷脂(DSPC/TPL)及饱和磷脂,总蛋白(DSPC/TP)、肺表面张力和白细胞计数,并行肺组织损伤评分。结果Ⅲ、Ⅳ、Ⅴ、Ⅵ组BALF中DSPC/TP及肺表面张力较Ⅰ组下降(P<0.05),通气结束时Crs较通气前下降(P<0.05),而Ⅱ组无显著性变化(P>0.05);与通气前比较,各组通气结束时MetHb与:NO2-/NO3-水平无变化,各组BALF中白细胞计数、肺组织损伤评分、肺泡扩张度和湿/干重比之间比较差异无统计学意义。结论1.3MAC异氟醚或七氟醚混合20ppmNO吸入可以安全用     

七氟醚介导腺苷A3受体通路减轻大鼠机械通气肺损伤

目的探讨腺苷A3受体(A_3AR)在七氟醚减轻大鼠机械通气肺损伤(VILI)中的作用及其机制。方法 SPF级健康雄性成年大鼠40只,体重200~250g,随机分为五组:假通气组(Sham组),模型对照组(CON组),七氟醚处理组(SEV组),七氟醚联合A_3AR抑制剂处理组(SM组),A_3AR抑制剂处理组(MRS组),每组8只。除Sham组不进行通气外,其他四组均在麻醉气管插管后行机械通气(V_T 12ml/kg),持续正压通气6h,建立大鼠VILI模型。Sham组:氯胺酮麻醉后直接处死;CON组:氯胺酮维持麻醉状态;SEV组:2%七氟醚维持麻醉;SM组:机械通气开始前30min腹腔注射MRS-1191 1mg/kg,2%七氟醚维持麻醉;MRS组:机械通气开始前30min腹腔注射MRS-1191 1mg/kg,氯胺酮维持麻醉。实验结束后处死大鼠,收集支气管肺泡灌洗液(BALF)和肺组织标本。采用Western blot法检测肺组织A_3AR、gp91~(phox)蛋白含量。检测BALF中炎性因子TNF-α、IL-1β浓度。DHE染色荧光检测肺组织活性氧(ROS)活性。观察肺组织病理学变化,并进行肺损伤评分。结果与SEV组比较,CON组、SM组和MRS组TNF-α、IL-β浓度明显升高(P0.05);与SM组比较,CON组与MRS组TNF-α、IL-β浓度明显升高(P0.05)。与Sham组比较,CON组、SEV组、SM组和MRS组A_3R、gp91~(phox)蛋白含量明显升高,CON组、SM组和MRS组ROS活性明显增强,肺损伤评分明显升高(P0.05);与CON组比较,SEV组和SM组A_3AR蛋白含量明显升高,SEV组gp91~(phox)蛋白含量、ROS活性和肺损伤评分明显降低(P0.05);与SEV组比较,SM组A_3AR蛋白含量明显降低,gp91~(phox)蛋白含量明显升高,SM组和MRS组ROS活性明显增强,肺损伤评分明显升高(P0.05)。结论七氟醚可以减轻机械通气诱发的肺损伤,其作用机制是通过A_3AR抑制肺脏氧化应激和炎症反应起效的。     

亚甲蓝对大鼠机械通气相关肺损伤中核苷酸结合寡聚化结构域样受体蛋白3炎性小体的影响

目的评价亚甲蓝对大鼠机械通气相关损伤(VILI)中核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎性小体的影响。方法清洁级成年雄性SD大鼠36只,8周龄,体重240～260 g,采用随机数字表法分为三组:自主呼吸对照组(C组)、大潮气量模型组(H组)和大潮气量+亚甲蓝组(MB组),每组12只。三组大鼠均行气管切开插管术,MB组经腹腔注射1%亚甲蓝50 mg/kg,10 min后以V_T 20 ml/kg行机械通气;C组经腹腔注射等容量生理盐水后保持自主呼吸;H组经腹腔注射等容量生理盐水,10 min后以V_T 20 ml/kg行机械通气。通气参数:FiO_2 21%,I∶E 1∶1,RR 80次/分,PEEP 0,通气时间4 h。于基础状态、通气结束时经颈总动脉取血行血气分析,通气结束后颈总动脉放血处死大鼠,收集肺组织标本、支气管肺泡灌洗液(BALF)。光镜下观察肺组织病理学改变,记录肺损伤评分,计算肺组织湿/干重比值(W/D)。采用ELISA法测定BALF中总蛋白含量以及血清、BALF中白细胞介素(IL)-1β、IL-18浓度;鲁米诺化学发光法检测肺组织中活性氧(ROS)含量;RT-PCR法及Western blot法分别检测肺组织NLRP3、凋亡相关斑点样蛋白(ASC)、天冬氨酸半胱氨酸蛋白酶-1(caspase-1)mRNA表达量及蛋白含量。结果与C组比较,H组肺损伤评分、W/D明显升高(P0.01),BALF中总蛋白以及血清、BALF中IL-1β和IL-18浓度明显升高(P0.01),肺组织中ROS含量、NLRP3、ASC、caspase-1 mRNA表达量及蛋白含量明显升高(P0.01)。与H组比较,MB组肺损伤评分、W/D明显降低(P0.05),BALF中总蛋白以及血清、BALF中IL-1β和IL-18浓度明显降低(P0.05),肺组织中ROS含量、NLRP3、ASC、caspase-1 mRNA表达量及蛋白含量明显降低(P0.05)。结论亚甲蓝通过抑制大鼠肺组织中NLRP3炎性小体的激活,阻碍促炎因子IL-1β及IL-18的形成,减轻大鼠VILI。     

外源性肺泡表面活性物质对大鼠呼吸机相关性肺损伤的保护作用

目的探讨气管注入肺泡表面活性物质(PS)对大鼠呼吸机相关性肺损伤(VILI)的保护作用及机制。方法将40只Wistar大鼠采用区组法随机分为对照组、高潮气量组(HV组)、VILI组和PS组4组,每组各10只。对照组未行机械通气,其余3组容量控制通气,PS组同时经气管内插管注入猪PS100mg/kg。监测心率、平均动脉压(MAP)、血气分析、肺湿/干重比(W/D)和支气管肺泡灌洗液(BALF)中白细胞计数,测定肺组织核因子-κB(NF-κB)活性和BALF中及血清白细胞介素-8(IL-8)浓度,并观察大体及光学显微镜下肺组织损伤的改变。结果损伤性机械通气后大鼠MAP及氧合指数(PaO2/FiO2)显著降低,而肺组织NF-κB活性和W/D显著增高。PS组与VILI组相比,MAP、PaO2/FiO2、肺组织NF-κB活性、BALF中白细胞计数、BALF及血清中IL-8浓度均有统计学意义(P<0.05)。组织学检查显示PS组较VILI组病变明显减轻。结论经气管注入PS可抑制VILI大鼠NF-κB基因表达,对VILI有保护作用。     

七氟醚对内毒素致急性肺损伤鼠肺泡毛细血管膜通透性的影响

目的　探讨七氟醚对内毒素致急性肺损伤鼠肺泡毛细血管膜通透性和肺泡灌洗液内炎性细胞的影响。方法　 4 8只Wistar大鼠 ,麻醉后静注伊万斯蓝 5 0mg/kg后 ,随机分为 4组 ,每组 12只。对照组 :股静脉注射生理盐水 1 2ml后机械通气 4h ;内毒素组 :股静脉注射内毒素 5mg/kg后机械通气 4h ;七氟醚 1L组和七氟醚 2L组 :股静脉注射内毒素 5mg/kg后机械通气 ,分别吸入肺泡气最低有效浓度 (MAC)为 1 0和 1 5的七氟醚 4h。 4h后取肺组织测定 :病理形态学积分 ,肺湿 /干重比 ,肺水含量 ,肺通透指数 ,伊万斯蓝含量和肺泡灌洗液内炎性细胞总数及百分比。结果　七氟醚1L组肺通透指数、伊万斯蓝含量、病理形态学积分分别由 4 6 8± 0 82 ,( 112 2 1± 11 4 4 )ng/mg ,9 17± 0 90下降到 3 98± 0 5 0 ,( 92 85± 11 80 )ng/mg ,7 5 0± 0 96 ;七氟醚 2L组下降到 3 91±0 34,( 96 33± 8 79)ng/mg ,7 6 7± 0 75。结论　 1 0和 1 5MAC七氟醚可降低内毒素所致急性肺损伤肺泡毛细血管膜通透性 ,使肺组织病理损伤减轻     

七氟醚预处理对大鼠单肺通气时肺组织血红素氧合酶-1表达的影响

目的 评价七氟醚预处理对大鼠单肺机械通气时肺组织血红素氧合酶-1(HO-1)表达的影响.方法 健康雄性SD大鼠24只,体重220～250 g,采用随机数字表法,将其随机分为4组(n=6):对照组(C组)、双肺通气组(T组)、单肺通气组(O组)和七氟醚预处理+单肺通气组(SO组).T组气管插管后行双肺通气1h,潮气量10ml/kg,通气频率60次/min,吸呼比1∶2,维持PETCO2 35～50 mmHg;O组和SO组气管插管后行右侧单肺通气1h,潮气量5ml/kg,通气频率80次/min,吸呼比1∶2,维持PETCO2 35～50 mm Hg.SO组单肺通气前吸入2.4％七氟醚30 min,纯氧洗脱15 min.C组和T组取左侧肺组织,O组和SO组取双侧肺组织,观察病理学结果,测定湿/干重比(W/D比)和HO-1表达.结果 与C组比较,T组左肺组织、O组和SO组双肺组织W/D比升高,HO-1表达上调(P＜0.05);与T组比较,O组双肺组织和SO组右肺组织W/D比升高,O组右肺组织和SO组双肺组织HO-1表达上调(P＜0.05);与O组比较,SO组双肺组织W/D比降低,HO-1表达上调(P＜0.05),病理学损伤减轻.结论 七氟醚预处理减轻大鼠单肺通气所致肺损伤的机制与其上调肺组织HO-1的表达有关.     

HSF1在小鼠呼吸机相关性肺损伤内源性保护机制中的作用：与HMGB1的关系

目的评价热休克转录因子1(HSF1)在小鼠呼吸机相关性肺损伤(VILI)内源性保护机制中的作用及其与高迁移率族蛋白B1(HMGB1)的关系。方法 SPF级健康C57BL/6雄性小鼠40只, 6～8周龄, 体质量20～25 g, 采用随机数字表法分为4组(n=10):对照组(C组)、VILI组、阴性对照siRNA+VILI组(NV组)和HSF1 siRNA+VILI组(siRNA组)。机械通气前48 h时, NV组气道内注射阴性对照siRNA 5 nmol, siRNA组气道内注射HSF1 siRNA 5 nmol, 均用无菌磷酸盐缓冲液稀释至50 μl。VILI组、NV组和siRNA组机械通气(VT 35 ml/kg, RR 75次/min, I∶E 1∶2, FiO2 21%)4 h构建小鼠VILI模型, C组仅行气管切开, 保留自主呼吸。分别于气管插管后即刻和机械通气4 h时, 行动脉血气分析, 记录PaO2;然后深麻醉下处死小鼠, 收集支气管肺泡灌洗液(BALF)和肺组织, 采用ELISA法测定BALF中TNF-α、IL-1β、HMGB1浓度, 测定肺组织湿重/干重(W/D)比值...     

基于慢病毒的SHARP-2特异的RNA干扰延长肾移植受者大鼠存活时间

Objective To investigate if rat enhancer of split- and hairy-related protein-2 (SHARP-2) short hairpin RNA interference (shRNAi) prolongs the survival time of rat kidney transplant recipients. Methods Gene recombinant procedures, transfection and co-transfection were carried out to introduce short hairpin RNA interference sequences target for SHARP-2 into 3rd generation self-inactivated lentiviral-ViraPower packaging mix. Limiting dilution method was used for viral titration. Real-time PCR was employed for quantification of gene expression. Rat kidney transplantation was utilized to investigate the effect of SHARP-2 gene silence on the recipient survival. Results A lentiviral-based shRNAi construct LV-SHARP-2iC showed 84% SHARP-2 gene silence efficiency in normal rat kidney cells. At multiplicity of infection 20, 57% T cells could be transfected by lentivirus with spinoculation method. In activated T cells, SHARP-2 g ene silence resulted in 61.3% and 68.7% reduction of intedeukin 2 (IL-2) and interferon γ (IFN-γ) gene expression. When donor kidney was perfused with 5×107 TU LV-SHARP-2iC, the median survival time prolonged for 4-5 days as compared to blank and scramble control groups. Conclusions A recombinant lentivirus LV-SHARP-2iC that effectively silence SHARP-2 gene expression is constructed successfully, leading to the inhibition of IL-2 and IFN-γ. LV-SHARP-2iC treatment can prolong the survival time of rat kidney transplant recipients.     

基于慢病毒的SHARP-2特异的RNA干扰延长肾移植受者大鼠存活时间

Objective To investigate if rat enhancer of split- and hairy-related protein-2 (SHARP-2) short hairpin RNA interference (shRNAi) prolongs the survival time of rat kidney transplant recipients. Methods Gene recombinant procedures, transfection and co-transfection were carried out to introduce short hairpin RNA interference sequences target for SHARP-2 into 3rd generation self-inactivated lentiviral-ViraPower packaging mix. Limiting dilution method was used for viral titration. Real-time PCR was employed for quantification of gene expression. Rat kidney transplantation was utilized to investigate the effect of SHARP-2 gene silence on the recipient survival. Results A lentiviral-based shRNAi construct LV-SHARP-2iC showed 84% SHARP-2 gene silence efficiency in normal rat kidney cells. At multiplicity of infection 20, 57% T cells could be transfected by lentivirus with spinoculation method. In activated T cells, SHARP-2 g ene silence resulted in 61.3% and 68.7% reduction of intedeukin 2 (IL-2) and interferon γ (IFN-γ) gene expression. When donor kidney was perfused with 5×107 TU LV-SHARP-2iC, the median survival time prolonged for 4-5 days as compared to blank and scramble control groups. Conclusions A recombinant lentivirus LV-SHARP-2iC that effectively silence SHARP-2 gene expression is constructed successfully, leading to the inhibition of IL-2 and IFN-γ. LV-SHARP-2iC treatment can prolong the survival time of rat kidney transplant recipients.     

基于慢病毒的SHARP-2特异的RNA干扰延长肾移植受者大鼠存活时间

Objective To investigate if rat enhancer of split- and hairy-related protein-2 (SHARP-2) short hairpin RNA interference (shRNAi) prolongs the survival time of rat kidney transplant recipients. Methods Gene recombinant procedures, transfection and co-transfection were carried out to introduce short hairpin RNA interference sequences target for SHARP-2 into 3rd generation self-inactivated lentiviral-ViraPower packaging mix. Limiting dilution method was used for viral titration. Real-time PCR was employed for quantification of gene expression. Rat kidney transplantation was utilized to investigate the effect of SHARP-2 gene silence on the recipient survival. Results A lentiviral-based shRNAi construct LV-SHARP-2iC showed 84% SHARP-2 gene silence efficiency in normal rat kidney cells. At multiplicity of infection 20, 57% T cells could be transfected by lentivirus with spinoculation method. In activated T cells, SHARP-2 g ene silence resulted in 61.3% and 68.7% reduction of intedeukin 2 (IL-2) and interferon γ (IFN-γ) gene expression. When donor kidney was perfused with 5×107 TU LV-SHARP-2iC, the median survival time prolonged for 4-5 days as compared to blank and scramble control groups. Conclusions A recombinant lentivirus LV-SHARP-2iC that effectively silence SHARP-2 gene expression is constructed successfully, leading to the inhibition of IL-2 and IFN-γ. LV-SHARP-2iC treatment can prolong the survival time of rat kidney transplant recipients.     

基于慢病毒的SHARP-2特异的RNA干扰延长肾移植受者大鼠存活时间

Objective To investigate if rat enhancer of split- and hairy-related protein-2 (SHARP-2) short hairpin RNA interference (shRNAi) prolongs the survival time of rat kidney transplant recipients. Methods Gene recombinant procedures, transfection and co-transfection were carried out to introduce short hairpin RNA interference sequences target for SHARP-2 into 3rd generation self-inactivated lentiviral-ViraPower packaging mix. Limiting dilution method was used for viral titration. Real-time PCR was employed for quantification of gene expression. Rat kidney transplantation was utilized to investigate the effect of SHARP-2 gene silence on the recipient survival. Results A lentiviral-based shRNAi construct LV-SHARP-2iC showed 84% SHARP-2 gene silence efficiency in normal rat kidney cells. At multiplicity of infection 20, 57% T cells could be transfected by lentivirus with spinoculation method. In activated T cells, SHARP-2 g ene silence resulted in 61.3% and 68.7% reduction of intedeukin 2 (IL-2) and interferon γ (IFN-γ) gene expression. When donor kidney was perfused with 5×107 TU LV-SHARP-2iC, the median survival time prolonged for 4-5 days as compared to blank and scramble control groups. Conclusions A recombinant lentivirus LV-SHARP-2iC that effectively silence SHARP-2 gene expression is constructed successfully, leading to the inhibition of IL-2 and IFN-γ. LV-SHARP-2iC treatment can prolong the survival time of rat kidney transplant recipients.     

直肠癌远端移形黏膜COX-2及BCL-2蛋白的表达与临床意义

目的探讨直肠癌远端移形黏膜COX-2及BCL-2蛋白的表达情况,判断直肠癌远端移形黏膜是否为癌前病变。方法应用高铁二胺-阿辛蓝染色检测54例直肠癌远端2cm处黏膜．将远端黏膜分为移形黏膜（TM）组及非移形黏膜（NTM）组,通过免疫组织化学染色检测TM中COX-2和BCL-2蛋白的表达．比较TM与NTM、肿瘤组织以及正常黏膜组织（20例直肠良性息肉旁肠黏膜组织）内BCL-2以及COX-2的表达的差异。结果54例直肠癌远端2cm黏膜处组织中有19例存在TM．35例为NTM。COX-2蛋白在肿瘤组织、TM、NTM、正常组织中的阳性率分别为81．5％（44／54）、21．1％（4／19）、17．1％（6／35）、10．0％（2／20）;BCL-2蛋白在上述4种组织中的阳性率分别为77．8％（42／54）、21．1％（4／19）、22．9％（8／35）、5．0％（1／20）。TM内的COX-2及BCL．2蛋白的表达与肿瘤组织相比,差异有统计学意义[（0．737±0．895）比（3．519±1．998）;（0．632±0．955）比（2．833±1．756）,均P〈0．01];与NTM、正常肠黏膜组织相比,差异无统计学意义（均P〉0．05）。结论直肠癌远端TM内的COX-2和BCL-2蛋白的表达无特异性．TM是癌前病变的证据不足。     

自由基清除剂对创伤后异位骨化形成中BMP-2及COX-2表达的影响

目的 研究自由基清除剂依达拉奉对创伤后异位骨化(HO)形成中骨形态发生蛋白(BMP)-2及环氧化酶(COX)-2表达的调节作用.方法 54只新西兰白兔随机分成实验组、模型组、对照组,每组18只.实验组和模型组兔适应性喂养1周后建立HO模型,对照组兔不建立HO模型,操作后第1天即开始给药.实验组给予依达拉奉30mg/次,...     

熊果酸对肝癌细胞株Bel-7404转移及侵袭能力的影响

目的 观察熊果酸(UA)对肝癌细胞株Bel-7404细胞转移侵袭能力及相关基因基质金属蛋白酶(MMP)-2、金属蛋白酶组织抑制因子(TIMP)-2 mRNA和蛋白表达水平的影响.方法 选用3个组:A组(UA 50 μmol/L)、B组(顺铂10mg/L)和C组(DMEM空白对照)对Bel-7404细胞进行处理,用细胞迁移实验及Transwell小室法检测细胞的迁移及侵袭能力,逆转录-聚合酶链反应(RT-PCR)及Western blot法检测细胞中MMP-2、TIMP-2 mRNA及蛋白的表达水平.结果 对Bel-7404细胞作用48 h后,A、B两组细胞的迁移速率(35.2±8.7)、(30.5±8.6)μm/h及侵袭穿膜细胞数(21.2±5.3)、(20.2±5.7)个、MMP-2 mRNA 0.24±0.06、0.23±0.05及蛋白0.21±0.01、0.24±0.04表达水平均较C组(75.6±8.7)μm/h、(54.8±7.8)个、0.46±0.11、0.42±0.06明显降低,差异均有统计学意义(P<0.01),而TIMP-2 mRNA0.87±0.05、0.83±0.06及蛋白0.98±0.06、0.95±0.09表达水平均比C组0.31±0.02、0.59±0.02高,差异均有统计学意义(P<0.01);A、B两组间细胞的迁移速率、侵袭穿膜细胞数、MMP-2和TIMP-2 mRNA及蛋白表达水平的比较差异均无统计学意义(P>0.05).结论 UA可抑制肝癌Bel-7404细胞株的迁移侵袭,其机制可能与MMP-2表达下调而TIMP-2表达上调有关.UA 50 μmol/L对Bel-7404细胞迁移侵袭能力的抑制作用与顺铂10 mg/L具有相同的效果及机制.     

基于慢病毒的SHARP-2特异的RNA干扰延长肾移植受者大鼠存活时间

Objective To investigate if rat enhancer of split- and hairy-related protein-2 (SHARP-2) short hairpin RNA interference (shRNAi) prolongs the survival time of rat kidney transplant recipients. Methods Gene recombinant procedures, transfection and co-transfection were carried out to introduce short hairpin RNA interference sequences target for SHARP-2 into 3rd generation self-inactivated lentiviral-ViraPower packaging mix. Limiting dilution method was used for viral titration. Real-time PCR was employed for quantification of gene expression. Rat kidney transplantation was utilized to investigate the effect of SHARP-2 gene silence on the recipient survival. Results A lentiviral-based shRNAi construct LV-SHARP-2iC showed 84% SHARP-2 gene silence efficiency in normal rat kidney cells. At multiplicity of infection 20, 57% T cells could be transfected by lentivirus with spinoculation method. In activated T cells, SHARP-2 g ene silence resulted in 61.3% and 68.7% reduction of intedeukin 2 (IL-2) and interferon γ (IFN-γ) gene expression. When donor kidney was perfused with 5×107 TU LV-SHARP-2iC, the median survival time prolonged for 4-5 days as compared to blank and scramble control groups. Conclusions A recombinant lentivirus LV-SHARP-2iC that effectively silence SHARP-2 gene expression is constructed successfully, leading to the inhibition of IL-2 and IFN-γ. LV-SHARP-2iC treatment can prolong the survival time of rat kidney transplant recipients.     

基于慢病毒的SHARP-2特异的RNA干扰延长肾移植受者大鼠存活时间

Objective To investigate if rat enhancer of split- and hairy-related protein-2 (SHARP-2) short hairpin RNA interference (shRNAi) prolongs the survival time of rat kidney transplant recipients. Methods Gene recombinant procedures, transfection and co-transfection were carried out to introduce short hairpin RNA interference sequences target for SHARP-2 into 3rd generation self-inactivated lentiviral-ViraPower packaging mix. Limiting dilution method was used for viral titration. Real-time PCR was employed for quantification of gene expression. Rat kidney transplantation was utilized to investigate the effect of SHARP-2 gene silence on the recipient survival. Results A lentiviral-based shRNAi construct LV-SHARP-2iC showed 84% SHARP-2 gene silence efficiency in normal rat kidney cells. At multiplicity of infection 20, 57% T cells could be transfected by lentivirus with spinoculation method. In activated T cells, SHARP-2 g ene silence resulted in 61.3% and 68.7% reduction of intedeukin 2 (IL-2) and interferon γ (IFN-γ) gene expression. When donor kidney was perfused with 5×107 TU LV-SHARP-2iC, the median survival time prolonged for 4-5 days as compared to blank and scramble control groups. Conclusions A recombinant lentivirus LV-SHARP-2iC that effectively silence SHARP-2 gene expression is constructed successfully, leading to the inhibition of IL-2 and IFN-γ. LV-SHARP-2iC treatment can prolong the survival time of rat kidney transplant recipients.     

地氟醚维持麻醉对SjvO2和CSFP的影响

研究地氟醚维持麻醉时ＰａＣＯ２对ＳｊｖＯ２和ＣＳＦＰ的影响。方法：４３例脑肿瘤病人用地氟醚维持麻醉,术中持续监测颅内压和动脉压。当人为改变ＰＥＴＣＯ２时,取颈内静脉血和同步采集动脉血作血气分析和测定两者乳酸含量。