老年病人冠脉搭桥术中脑氧代谢与术后精神障碍的关系

目的 观察老年病人体外循环(CPB)冠脉搭桥术(CABG)中脑氧代谢变化与术后精神障碍的关系。方法 年龄≥65岁择期CABG病人30例,取右颈内静脉和桡动脉血进行血气分析,葡萄糖及乳酸测定,并计算脑血流量/脑氧耗比值(CBF/CMRO_2)、脑氧耗/脑糖耗比值(CMRO_2/CMRglu)及乳酸生成量(ADVL);术后用ICU精神错乱评估量表(CAM-ICU)将病人分为精神障碍(POMD)组和无精神障碍(NPMD)组,比较两组术中指标差异。结果 (1)术后7例病人发生POMD,发生率为23.33％;(2)复温期8例病人颈内静脉血氧发生去饱和(颈内静脉血氧饱和度≤50％或颈内静脉血氧分压≤25mmHg),其中POMD组3例(发生率43％),NPMD组5例(发生率22％),两组差异显著(P<0.01);(3)复温期POMD组CBF/CMRO_2显著低于NPMD组(P<0.01);(4)降温期、复温期和停CPB,尤其是复温期,POMD组CMRO_2/CMRglc明显低于NPMD组,P<0.01,而ADVL明显高于后者(P<0.01)。结论老年病人CABG术后精神障碍与CPB期间脑氧代谢失衡有关。     

深低温低流量灌注与降温期血气管理对乳猪脑保护的对照研究

目的：在深低渐体外循环中比较降温期pH稳态血气管理和深低温阶段低流量灌注对脑保护的作用能力。方法：24头乳猪根据不同的体外循环脑保护处理方法分成4组，A组：深低温停循环、降温期alpha稳态血气管理；B组：深低温停循环、降温期pH稳态血气管理；C组：深低温低流量、降温期alpha稳态血气管理；D组：深低温低流量、降温期pH稳态血气管理。比较不同脑保护方法和体外循环阶段对脑血流（CBF）、脑氧代谢率（CMRO2）、脑乳酸含量、脑水含量和脑电图（EEG）的影响。结果：B组、D组降温末CBF均高于A组、C组；而CMRO2则低于A组、C组；B组、D组降温期EEC平坦波出现早。C组、D组复温期CBF、CMRO2和EEG恢复好于A组、B组。复温末A组、B组颈内静脉乳酸含量显著高于C组、D组，脑水含量组间差异无显著性。结论：深低温体外循环流量灌注对脑保护的作用能力强于降温期pH稳态血气管理，两种方法配合应用具有脑保护的协同作用。     

急性高容血液稀释对老年心脏手术病人脑氧合功能的影响

目的　观察急性高容性血液稀释 (AHHD)对老年心脏手术病人脑氧合功能的影响并探讨其可能的作用机制。方法　 2 4例冠状动脉搭桥术 (CABG)的病人 ,ASAⅠ～Ⅱ级 ,随机分为老年扩容组、未扩容组和中青年扩容组。扩容组诱导时 1小时内经颈内静脉推注 6 %贺斯溶液 1 0ml/kg ,同时静注硝酸甘油 0 5μg·kg- 1 ·min- 1 。分别于术前、诱导后 5分钟、扩容止及主动脉插管时监测血液动力学指标 ,测定动、静脉血乳酸浓度 ,脑血氧饱和度 (SjvO2 ) ,脑血氧分压 (PjO2 )和脑血流速率。结果　 (1 )老年扩容组的PAP较术前显著增高 (P <0 0 5) ,但与其他两组相比无显著差异。 (2 )未扩容组的SjvO2 ,CBF/CMRO2 在主动脉插管时较术前显著降低 (P <0 0 1 ) ,与扩容组相比也显著减少 (P<0 0 5) ;而老年扩容组SjvO2 较术前及另外两组均显著增加 (P <0 0 5) ,CBF/CMRO2 无明显变化。(3)脑血流速率在中青年扩容组病人均显著高于另外两组 (P <0 0 5)。 (4)脑乳酸净生成量 (ADVL)三组间无显著差异 (P >0 0 5)。结论　AHHD对老年手术病人脑氧合功能具有一定的保护作用。     

不同复温速率对体外循环下冠脉搭桥术患者脑氧代谢及脑电活动的影响

目的观察不同复温速率对体外循环(CPB)下冠脉搭桥术患者脑氧代谢及脑电活动的影响。方法择期CPB下行冠脉搭桥术患者30例,ASAⅡ或Ⅲ级,随机分为2组,快速复温组(A组,n =14):复温时间≤20 min;缓慢复温组(B组,n=16):复温时间>20 min。CPB亚低温时鼻咽温(NPT) 维持在26-27℃,当NPT达36.5-37.50℃时复温结束,分别于CPB前、复温前5 min、复温开始后每隔5 min至CPB后5 min各时点记录NPT、平均动脉压(MAP)、脑电双频指数(BIS),并取桡动脉及颈内静脉血,进行血气分析,复温过程中每两个观察时点的温度变化速率(△dT)与颈内静脉血氧饱和度(SjvO2)差值(△SjvO2)进行直线回归分析,NPT分别与SjvO2、动脉-颈静脉血氧含量差[C(a-ja)O2]、BIS进行Spearman相关分析;记录术后神经系统并发症情况。结果随着温度的升高,A组SjvO2、颈内静脉血氧分压下降,C(a-j)O2上升,B组上述指标差异无统计学意义(P>0.05),两组BIS升高(P<0.05或0.01)。NPT与SjvO2、C(a-jv)O2、BIS的Spearman相关系数分别-0.524、0.642、0.848(P<0.01)。△dT (X)与△SjvO2(Y)的直线回归方程为lnY=ln12.65 1.31lnX,拟合的最优曲线为Power幂曲线模型。A组有2例出现术后神经系统并发症,B组均未出现术后神经系统并发症。结论CPB下冠脉搭桥术患者缓慢复温可降低脑电活动和脑代谢率,减少脑损伤。     

心肺转流时利多卡因对脑氧代谢的影响

目的　观察心肺转流 (CPB)时利多卡因对脑氧代谢的影响,探讨脑保护措施。方法　二尖瓣膜置换手术20例,随机均分为对照组 (Ⅰ组)和利多卡因组(Ⅱ组)。Ⅱ组在麻醉后用微量泵输入利多卡因150μg·kg 1·min 1。α稳态处理血气,通过桡动脉和颈内静脉球部血液的动态血气分析及乳酸浓度测定,计算全身动脉及颈内静脉的血氧含量、动 颈内静脉血氧含量差、脑氧摄取率和动 颈内静脉血乳酸浓度差。分析CPB时利多卡因对脑氧代谢的影响。结果　CPB开始后,两组的动脉血氧含量、动 颈内静脉血氧含量差和脑氧摄取率下降,随着复温又升高。CPB降温至 26℃时,Ⅱ组的动 颈内静脉血氧含量差、脑氧摄取率明显低于Ⅰ组(P<0 .01)。两组的乳酸浓度均升高,降温后动 颈内静脉血的乳酸浓度差无明显变化。停CPB时,两组的动 颈内静脉血氧含量差仍明显低于CPB前水平(P<0 .01)。结论　CPB中利多卡因能降低脑氧耗,有利于改善大脑氧供需平衡和保护CPB中缺血脑细胞的功能。     

星状神经节阻滞对心脏瓣膜置换术患者脑氧代谢的影响

目的 观察星状神经节阻滞(SGB)对心肺转流(CPB)下心脏瓣膜置换术患者脑氧代谢的影响.方法 择期行心脏瓣膜置换术患者40例,随机均分为SGB组和对照组.SGB组以0.25%罗哌卡因10 ml行右侧SGB,出现Horner综合征为有效;对照组不行SGB.左颈内静脉逆行穿刺置管至颈静脉球部备采集血标本,分别于左颈内静脉逆行置管后即刻(T0)、降温至鼻咽温32℃(T1)、CPB后30 min(T2)、复温至鼻咽温36℃(T3)、停CPB时(T4)抽取颈内静脉球部血和桡动脉血做血气分析.并检测乳酸和血糖,同时取颈内静脉球部血5 ml检测血浆内皮素-1(ET-1)、降钙素基因相关肽(CGRP)浓度,按Fick公式计算动脉血氧含量(CaO2)、颈内静脉血氧含量(CjvO2)、动静脉血氧含量差(Ca-vO2)、脑氧摄取率(O2ER)、桡动脉-颈内静脉乳酸差值(ADVL)及桡动脉-颈内静脉血糖差值(Ga-v).结果 SGB组ET-1在T2～T4时明显低于对照组(P<0.01),T3、T4时SGB组CGRP/ET-1比值高于对照组(P<0.05).T1～T4时SGB组SjvO2显著高于、CaO2、CjvO2、Ca-vO2、O2 ER及Ga-v显著低于T0时(P<0.05或P<0.01).对照组CaO2、CjvO2、Ca-vO2亦明显低于T0时(P<0.01).SGB组ADVL各时点差异无统计学意义.对照组ADVL在T3、T4时高于T0时和SGB组(P<0.05或P<0.01).结论 SGB可以调节CGRP/ET-1相对平衡,有利于改善脑组织灌注,从而改善CPB期间脑氧代谢平衡紊乱.     

体外循环术中低温改良低钾右旋糖酣肺动脉灌注的肺保护作用

目的　研究体外循环(cardiopulmanarybypass, CPB)期间低温改良低钾右旋糖酐(low patassiumdextran, LPD)溶液肺动脉灌注对心脏瓣膜置换病人的肺保护作用。　方法　30例行二尖瓣置换术病人随机分为两组:肺灌注组(15例),CPB术中一次性肺动脉灌注低温改良LPD液;对照组(15例)常规行二尖瓣置换术,未行灌注。分别于术前、CPB结束、手术结束、术后6h测算氧合指数(PaO2 /FiO2 )、肺静态顺应性变化。CPB停机后30min,取病人右上肺组织,观察组织形态学变化。　结果　肺灌注组病人CPB结束、手术结束、术后6h的氧合指数分别为(421±31)、(382±41)、(370±39)mmHg,肺静态顺应性分别为(30. 8±3. 6)、(29. 2±3. 3)、(29. 2±3. 1)ml/cmH2O,明显高于对照组的( 340±33), (321+38), (315±41)mmHg和(24. 2±3. 0)、( 21. 3±2. 8 )、( 19. 0±3. 0 )ml/cmH2O (P<0. 05 )。肺组织活检病理检查结果显示,对照组肺间质水肿明显,有大量炎性细胞浸润,肺灌注组无明显病理改变。　结论　CPB术后存在肺损伤,CPB中采用低温改良LPD液肺动脉灌注具有肺保护作用。     

体外循环期间血浆一氧化氮浓度变化

目的　吸入一氧化氮 (NO)应用于心内直视手术治疗肺动脉高压取得较好作用 ,感染性休克病人血浆NO浓度明显升高 ,但心肺转流 (CPB)期间血浆NO浓度变化报道尚少。本文测定了CPB期间及停机后血浆NO浓度的变化 ,旨在为临床应用NO治疗提供参考。方法　分别在手术前、CPB10分钟、停机前及停机后 1、6、12小时和 2 4小时测量血浆NO浓度。结果　术前血浆NO平均值 ( 2 73± 0 67) μmol/L ,而CPB后 10分钟、停机前及停机后 1、6、12小时和 2 4小时分别为 ( 3 80± 1 61)、( 4 0 9± 1 3 0 )、( 3 97± 1 18)、( 3 80± 1 3 1)、( 3 69± 1 0 2 )和 ( 3 4 1± 1 2 1) μmol/L。停机前达最高值 ;停机后浓度有所降低 ,但均较术前值显著升高 (P <0 0 0 0 1) ;术后 2 4小时仍高于术前。结论　CPB可导致血浆NO浓度显著升高 ,且持续至停止CPB后 2 4小时。CPB降温和复温过程对NO有影响 ,复温期NO浓度达最高值。NO浓度升高可能与CPB过程中的全身炎症反应有关。     

孕羊深低温体外循环对胎羊温度和血流动力学及血气的影响

目的探讨孕羊深低温体外循环(CPB)对胎羊温度、血流动力学和血气的影响。方法5头健康怀孕山羊常规建立CPB,转流降温、复温各1 h。监测孕羊和胎羊的温度、心率、平均动脉压、血气值。结果孕羊最低温度(17．4±1．5)℃,胎羊最低温度(24．6±1．5)℃。降温期胎羊温度始终高于孕羊温度,复温期孕羊-胎羊温差逆转,转流结束胎羊温度低于孕羊温度。降温期胎羊心率逐渐减慢,复温期不能恢复正常心率。低温转流15 min,胎羊pH值从转流开始的7．30± 0．03降到7．17±0．07(P<0．05)、PO_2从转流开始的(32．5±4．0)mmHg(1 mmHg=0．133 kPa)降到(17．5±3．0)mm Hg(P<0．05),PCO_2从转流开始的(44．8±2．2)mm Hg升到(56．8±5．1)mm Hg(P<0．05),BE值从转流开始的(-3．2±0．6)升到(-5．7±1．3)(P<0．05),此后血气值进一步恶化,复温阶段胎羊血气值也没有好转。CPB结束胎羊存活率为60％。结论孕羊深低温CPB 影响胎盘的热交换和气体交换功能,对胎羊存活不利。     

异氟烷和异丙酚对体外循环心内直视手术患者围术期脑氧代谢和脑损伤影响的研究

目的：观察异氟烷和异丙酚麻醉对体外循环（CPB）心内直视手术患者围术期脑氧供需平衡及脑损伤标记物S100β和神经元性烯醇化酶（NSE）蛋白浓度的影响。方法：将30例择期行体外循环瓣膜置换手术病人随机分为异氟烷麻醉组和异丙酚麻醉组。分别于CBP开始前（T1）、鼻咽温降至稳定期（L）．复温至36℃（T3）,CBP结束后30m;n（T4）．CPB结束后6h（T5）和CPB结束后24h（T6）抽取桡动脉血和颈静脉球部血样检测血气并测定颈内静脉血S-100B和NSE蛋白的浓度。结果：CPB前两组患者颈内静脉球氧饱和度（Sjv02）,动脉一颈内静脉血氧含量差（Da—jv02）及氧摄取率（CERO2）和脑损害标记物s100B及NSE蛋白浓度差异无统计学意义（P〉0．05）;随着降温逐渐加深两组Sjv02明显升高．Da—jvO2和CERO2明显降低．复温开始后两组SjvO2较CPB前明显降低,而Da—jvO2．CERO2则明显升高（P〈0．05）;CPB开始后两组血浆s100β和NSE蛋白浓度逐渐升高。复温期达到峰值．s100β蛋白浓度在停机24h后恢复至基础水平,NsE蛋白浓度仍略高于CPB前水平;两组各指标比较．异丙酚组的变化幅度要明显低于异氟烷组。结论：两种麻醉方式均可满足CPB下瓣膜置换手术的需要,与异氟烷组比较．异丙酚麻醉更有利于CBP下患者脑氧代谢的改善和脑损伤的保护。     

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.     

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.     

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.