移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

Objective To investigate the expression and significance of local Th1 and Th2 cytokines in the grafts of pig simultaneous pancreas-kidney with portal versus systemic venous drainage. Methods Fifty-eight non-related and first hybrid landraces were divided into Sham (n = 10), PE (portal-enteric drainage) (n =24) and SE ( systemic-enteric drainage) (n =24) groups. Type 1 diabetic mellitus was induced by whole pancreatectomy ,and right-side nephrectomy was also performed in the recipients. The pigs in sham group were only subjected to ]aparotomy. The portal vein was anastomosed to superior mesenteric vein of the recipient in PE group or to the inferior hepatic cava of the recipients in SE group. The tissues of the graft pancreas and kidney were obtained by surgery for detecting the expression of mRNA and protein of IFN-γand IL4 by RT-PCR and ELISA on the day 3,and 7 after transplantation. Results Compared with sham group,the local expression of mRNA and protein of IFN in the graft pancreas and kidney of both SE and PE groups was significantly up-regulated ( P < 0. 05 ) ( On the day 7, protein expression in sham, SE, and PE groups :in the graft pancreas, [ (3.36±0.25 ), (7.72±0.67 ), and ( 6.52±0.34 ) pg/mg protein ];in the graft kidney, [ ( 3.66±0.28 ), ( 10.83±1.48 ), and (7.79±0.60) pg/mg protein ]. The local expression of mRNA and protein of IL-4 was significantly up-regulated in PE group (P < 0.05 ), but down-regulated or no expression was detected in group SE (on the day 7, protein expression in sham, SE, and PE groups:in the graft pancreas, [ (7.18±0.16), (6.10±0.16) ,and (20.66±1.47) pg/mg protein];in the graft kidney, [ (5.74±0.48) ,(10.38±0.92) ,and ( 19.66±1.57) pg/mg protein]. Conclusion That the blood flow of graft pancreas passes by liver in SPK with PVD can induce a Th1 to Th2 inmaune shift. This may be one of mechanisms of immunologic benefits of PVD.     

移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

Objective To investigate the expression and significance of local Th1 and Th2 cytokines in the grafts of pig simultaneous pancreas-kidney with portal versus systemic venous drainage. Methods Fifty-eight non-related and first hybrid landraces were divided into Sham (n = 10), PE (portal-enteric drainage) (n =24) and SE ( systemic-enteric drainage) (n =24) groups. Type 1 diabetic mellitus was induced by whole pancreatectomy ,and right-side nephrectomy was also performed in the recipients. The pigs in sham group were only subjected to ]aparotomy. The portal vein was anastomosed to superior mesenteric vein of the recipient in PE group or to the inferior hepatic cava of the recipients in SE group. The tissues of the graft pancreas and kidney were obtained by surgery for detecting the expression of mRNA and protein of IFN-γand IL4 by RT-PCR and ELISA on the day 3,and 7 after transplantation. Results Compared with sham group,the local expression of mRNA and protein of IFN in the graft pancreas and kidney of both SE and PE groups was significantly up-regulated ( P < 0. 05 ) ( On the day 7, protein expression in sham, SE, and PE groups :in the graft pancreas, [ (3.36±0.25 ), (7.72±0.67 ), and ( 6.52±0.34 ) pg/mg protein ];in the graft kidney, [ ( 3.66±0.28 ), ( 10.83±1.48 ), and (7.79±0.60) pg/mg protein ]. The local expression of mRNA and protein of IL-4 was significantly up-regulated in PE group (P < 0.05 ), but down-regulated or no expression was detected in group SE (on the day 7, protein expression in sham, SE, and PE groups:in the graft pancreas, [ (7.18±0.16), (6.10±0.16) ,and (20.66±1.47) pg/mg protein];in the graft kidney, [ (5.74±0.48) ,(10.38±0.92) ,and ( 19.66±1.57) pg/mg protein]. Conclusion That the blood flow of graft pancreas passes by liver in SPK with PVD can induce a Th1 to Th2 inmaune shift. This may be one of mechanisms of immunologic benefits of PVD.     

猴猪非协调性异种胰肾联合移植的实验研究

目的: 尝试猴猪非协调性异种胰肾联合移植,观察异种移植间的免疫反应.方法: 整块切取猴的胰十二指肠、右肾及脾,保留带有腹腔动脉、肠系膜上动脉、右肾动脉的腹主动脉段.切除猪的胰腺制成I型糖尿病的模型,并切除猪的右肾.以端侧吻合的方式将供体猴腹主动脉段和门静脉与猪的肾后腹主动脉段和门静脉分别对应吻合;以端端吻合的方式将供受体的右肾静脉进行吻合;供体输尿管自腹壁引出腹腔.将猴的十二指肠与猪空肠行端侧吻合.结果: 移植物恢复灌注后5分钟开始出现超急性排斥反应,受体于术后19h 死亡.结论: 本实验是对异种移植研究的初探;猴→猪异种移植间的超急性排斥反应可能存在其它的机制,有待进一步研究.     

胆囊胆固醇结石病与载脂蛋白AⅠ—CⅢ—AⅣ基因PstⅠ、SstⅠ位点多态性关联的临床实验研究

目的：探讨载脂蛋白AⅠ—CⅢ—AⅣ基因簇多态性与胆囊胆固醇结石病的关系，揭示胆固醇结石病的遗传易感性。方法：应用多聚酶链反应技术（PCR）对161例胆固醇结石病患者及94例正常人载脂蛋白（Apo）AⅠ—CⅢ—AⅣ基因簇的限制性片段长度多态性（RFLP）进行研究，检测ApoAⅠ—CⅢ—AⅣ基因位点PstⅠ、SsfⅠ。结果：胆石组P1P1基因型和P1等位基因频率明显低于对照组，P1P2和P2B基因型以及P2等位基因频率明显高于对照组（P〈0．05），SsfⅠ酶切位点的基因型和等位基因频率在两组间的分布无显著性差异。结论：ApoAⅠ—CⅢ—AⅣ基因簇PsfⅠ位点的多态性可能与女性胆囊胆固醇结石病有关联。     

移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

Objective To investigate the expression and significance of local Th1 and Th2 cytokines in the grafts of pig simultaneous pancreas-kidney with portal versus systemic venous drainage. Methods Fifty-eight non-related and first hybrid landraces were divided into Sham (n = 10), PE (portal-enteric drainage) (n =24) and SE ( systemic-enteric drainage) (n =24) groups. Type 1 diabetic mellitus was induced by whole pancreatectomy ,and right-side nephrectomy was also performed in the recipients. The pigs in sham group were only subjected to ]aparotomy. The portal vein was anastomosed to superior mesenteric vein of the recipient in PE group or to the inferior hepatic cava of the recipients in SE group. The tissues of the graft pancreas and kidney were obtained by surgery for detecting the expression of mRNA and protein of IFN-γand IL4 by RT-PCR and ELISA on the day 3,and 7 after transplantation. Results Compared with sham group,the local expression of mRNA and protein of IFN in the graft pancreas and kidney of both SE and PE groups was significantly up-regulated ( P < 0. 05 ) ( On the day 7, protein expression in sham, SE, and PE groups :in the graft pancreas, [ (3.36±0.25 ), (7.72±0.67 ), and ( 6.52±0.34 ) pg/mg protein ];in the graft kidney, [ ( 3.66±0.28 ), ( 10.83±1.48 ), and (7.79±0.60) pg/mg protein ]. The local expression of mRNA and protein of IL-4 was significantly up-regulated in PE group (P < 0.05 ), but down-regulated or no expression was detected in group SE (on the day 7, protein expression in sham, SE, and PE groups:in the graft pancreas, [ (7.18±0.16), (6.10±0.16) ,and (20.66±1.47) pg/mg protein];in the graft kidney, [ (5.74±0.48) ,(10.38±0.92) ,and ( 19.66±1.57) pg/mg protein]. Conclusion That the blood flow of graft pancreas passes by liver in SPK with PVD can induce a Th1 to Th2 inmaune shift. This may be one of mechanisms of immunologic benefits of PVD.     

移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

Objective To investigate the expression and significance of local Th1 and Th2 cytokines in the grafts of pig simultaneous pancreas-kidney with portal versus systemic venous drainage. Methods Fifty-eight non-related and first hybrid landraces were divided into Sham (n = 10), PE (portal-enteric drainage) (n =24) and SE ( systemic-enteric drainage) (n =24) groups. Type 1 diabetic mellitus was induced by whole pancreatectomy ,and right-side nephrectomy was also performed in the recipients. The pigs in sham group were only subjected to ]aparotomy. The portal vein was anastomosed to superior mesenteric vein of the recipient in PE group or to the inferior hepatic cava of the recipients in SE group. The tissues of the graft pancreas and kidney were obtained by surgery for detecting the expression of mRNA and protein of IFN-γand IL4 by RT-PCR and ELISA on the day 3,and 7 after transplantation. Results Compared with sham group,the local expression of mRNA and protein of IFN in the graft pancreas and kidney of both SE and PE groups was significantly up-regulated ( P < 0. 05 ) ( On the day 7, protein expression in sham, SE, and PE groups :in the graft pancreas, [ (3.36±0.25 ), (7.72±0.67 ), and ( 6.52±0.34 ) pg/mg protein ];in the graft kidney, [ ( 3.66±0.28 ), ( 10.83±1.48 ), and (7.79±0.60) pg/mg protein ]. The local expression of mRNA and protein of IL-4 was significantly up-regulated in PE group (P < 0.05 ), but down-regulated or no expression was detected in group SE (on the day 7, protein expression in sham, SE, and PE groups:in the graft pancreas, [ (7.18±0.16), (6.10±0.16) ,and (20.66±1.47) pg/mg protein];in the graft kidney, [ (5.74±0.48) ,(10.38±0.92) ,and ( 19.66±1.57) pg/mg protein]. Conclusion That the blood flow of graft pancreas passes by liver in SPK with PVD can induce a Th1 to Th2 inmaune shift. This may be one of mechanisms of immunologic benefits of PVD.     

RNAi抑制胰腺癌细胞survivin表达并诱导细胞凋亡的实验研究

目的构建RNAi表达载体,并分析其对胰腺癌细胞PANC-1 survivin表达的抑制作用。方法用免疫荧光技术和RT-PCR检测胰腺癌细胞PANC-1中凋亡抑制蛋白survivin的表达,并把survivin基因克隆到T载体进行测序,构建针对survivin基因的干扰载体,用脂质体转染胰腺癌细胞PANC-1,RT-PCR、western-blot分析干扰载体对survivin表达的抑制情况,流式细胞仪检测细胞凋亡。结果survivin在胰腺癌细胞PANC-1中高表达,其基因序列与GenBank中公布的一致,干扰载体pTZU6＋1-svv2可以有效的抑制survivin的表达,抑制率约为74％,并诱导了肿瘤细胞的凋亡。结论用RNAi表达载体可以有效抑制胰腺癌细胞PANC-1中凋亡抑制蛋白survivin的表达并诱导胰腺癌细胞凋亡。     

移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

目的 探讨Th1和Th2型细胞因子在猪胰肾联合移植门脉(PVD)和体静脉(SVD)回流术式移植物局部表达及意义.方法 58例四川当地杂交第1代长白猪,分成假手术组(10例)、PE组(24例)和SE组(24例).假手术组行开腹术,PE组和SE组切除受者胰腺制成Ⅰ型糖尿病模型,并切除右肾.PE组门静脉与受者肠系膜上静脉吻合,SE组门静脉与受者肝下下腔静脉吻合,外分泌均采用肠道引流.术后第3、7天开腹取移植胰腺、肾脏组织,逆转录-聚合酶链反应(RT-PCR)和ELISA检测移植物局部Th1型细胞因子IFN-γ和Th2型细胞因子白细胞介素(IL)-4mRNA及蛋白的表达.结果 术后第3、7天,与对照组比较,移植肾和胰腺IFN-γ mRNA及蛋白在SE组和PE组均呈高表达(P<0.05),SE组高于PE组(P<0.05)(第7天,蛋白在对照组比SE组比PE组表达:移植胰腺,[(3.36±0.25)比(7.72±0.67)比(6.52±0.34)pg/mg蛋白],移植肾[(3.66±0.28)比(10.83±1.48)比(7.79±0.60)pg/mg蛋白].移植胰腺和肾IL-4 mRNA及蛋白在SE组和假手术组呈低表达,PE组呈高表达(P<0.05)(第7天,蛋白在对照组比SE组比PE组表达:移植胰腺,[(7.18±0.16)比(6.10±0.16)比(20.66±1.47)pg/mg蛋白];移植肾,[(5.74±0.48)比(10.38±0.92)比(19.66±1.57)pg/mg蛋白].结论 胰肾联合移植PVD术式移植物血流经过肝脏后可诱导,Th1细胞向Th2细胞偏离,是PVD免疫学优越性的可能机制.     

胆囊胆固醇结石病与载脂蛋白AI-CIII-AIV基因PstI、SstI位点多态性关联的临床实验研究

目的: 探讨载脂蛋白AI-CIII-AIV基因簇多态性与胆囊胆固醇结石病的关系,揭示胆固醇结石病的遗传易感性.方法: 应用多聚酶链反应技术(PCR)对161例胆固醇结石病患者及94例正常人载脂蛋白(Apo)AI-CIII-AIV基因簇的限制性片段长度多态性(RFLP)进行研究,检测ApoAI-CIII-AIV基因位点PstI、SstI.结果: 胆石组P1P1基因型和P1等位基因频率明显低于对照组,P1P2和P2P2基因型以及P2等位基因频率明显高于对照组(P＜0.05), SstI酶切位点的基因型和等位基因频率在两组间的分布无显著性差异.结论: ApoAI-CIII-AIV基因簇PstI位点的多态性可能与女性胆囊胆固醇结石病有关联.     

移植物局部Th1和Th2型细胞因子在猪胰肾联合移植门脉和体静脉回流术式的表达及其意义

Objective To investigate the expression and significance of local Th1 and Th2 cytokines in the grafts of pig simultaneous pancreas-kidney with portal versus systemic venous drainage. Methods Fifty-eight non-related and first hybrid landraces were divided into Sham (n = 10), PE (portal-enteric drainage) (n =24) and SE ( systemic-enteric drainage) (n =24) groups. Type 1 diabetic mellitus was induced by whole pancreatectomy ,and right-side nephrectomy was also performed in the recipients. The pigs in sham group were only subjected to ]aparotomy. The portal vein was anastomosed to superior mesenteric vein of the recipient in PE group or to the inferior hepatic cava of the recipients in SE group. The tissues of the graft pancreas and kidney were obtained by surgery for detecting the expression of mRNA and protein of IFN-γand IL4 by RT-PCR and ELISA on the day 3,and 7 after transplantation. Results Compared with sham group,the local expression of mRNA and protein of IFN in the graft pancreas and kidney of both SE and PE groups was significantly up-regulated ( P < 0. 05 ) ( On the day 7, protein expression in sham, SE, and PE groups :in the graft pancreas, [ (3.36±0.25 ), (7.72±0.67 ), and ( 6.52±0.34 ) pg/mg protein ];in the graft kidney, [ ( 3.66±0.28 ), ( 10.83±1.48 ), and (7.79±0.60) pg/mg protein ]. The local expression of mRNA and protein of IL-4 was significantly up-regulated in PE group (P < 0.05 ), but down-regulated or no expression was detected in group SE (on the day 7, protein expression in sham, SE, and PE groups:in the graft pancreas, [ (7.18±0.16), (6.10±0.16) ,and (20.66±1.47) pg/mg protein];in the graft kidney, [ (5.74±0.48) ,(10.38±0.92) ,and ( 19.66±1.57) pg/mg protein]. Conclusion That the blood flow of graft pancreas passes by liver in SPK with PVD can induce a Th1 to Th2 inmaune shift. This may be one of mechanisms of immunologic benefits of PVD.