胰腺导管腺癌(pancreatic ductal adenocarcinoma,PDAC)是恶性程度最高的肿瘤之一。越来越多的证据表明包括立体定向放射治疗(stereotactic body radiotherapy,STRT)和射频消融在内的局部治疗可能成为不可手术切除的PDAC患者的新选择。...胰腺导管腺癌(pancreatic ductal adenocarcinoma,PDAC)是恶性程度最高的肿瘤之一。越来越多的证据表明包括立体定向放射治疗(stereotactic body radiotherapy,STRT)和射频消融在内的局部治疗可能成为不可手术切除的PDAC患者的新选择。由于PDAC的单独免疫治疗效果欠佳,近年来越来越多的研究发现局部治疗可以改变肿瘤局部及远处转移灶的免疫微环境,并增加免疫治疗敏感性。可以预期,局部治疗联合免疫治疗可能提高疗效。本文主要介绍PDAC局部治疗联合免疫治疗的机制及相关临床研究进展。展开更多
AIM: To investigate the effects of electroporation on primaryrat hepatocyte and to optimize the electroporation conditionsintroducing foreign genes into primary hepatocytes.METHODS: A single-pulse procedure was perfor...AIM: To investigate the effects of electroporation on primaryrat hepatocyte and to optimize the electroporation conditionsintroducing foreign genes into primary hepatocytes.METHODS: A single-pulse procedure was performed at Iowvoltage (220-400 V) but with high capacitance (500-950 μF).Hepatocytes were divided into 4 groups according to theelectroporation conditions: group Ⅰ, 220 V and 500 μF;group Ⅱ, 220 Vand 950 μF; group Ⅲ, 400 V and 950 μF,and group Ⅳ.The control group was freshly isolatedhepatocytes and directly cultured under the same conditionsas those of electroporation groups. The effects ofelectroporation on primary rat hepatocytes were detectedby trypan blue exclusion (TBE) and MTT analysis. Besides,albumin (AIb), alanine transaminase (ALT) and lactatedehydrogenase (LDH) in the supernatants of culturedhepatocytes were measured by biochemical assay.RESULTS: Between day 1 and day 15 after incubation,primary rat hepatocytes of each electroporation groupappeared normal, being the same with those of controlgroup. TBE staining showed that slight hepatocyte damageand high survival rate were found in the electroporationgroups and the control group. Cultured for 3, 7, 11 and 15days, hepatocyte viability was approximatly 92.6±2.5 %,89.5±3.3 %, 82.0±3.5 % and 74.3±1.2 %, respectively.MTT analysis indicated that the viabilities of hepatocyteshad no significant difference between each electroporationgroup, and those were similar to that of control group. Atthe 36th hour after electroporation, AIb, ALT and LDH in thesupernatants of control group were 5.3±0.1 g. L-1, 183.7±8.4 nkat. L-1 and 896.8±58.5 nkat. L-1; those of group Ⅱwere 5.7±0.1 g. L-1, 215.4±16.7 nkat. L-1 and 1063.8±51.8 nkat. L-1; and those of group Ⅲ were 5.80.2 g. L-1,217.1 ± 8.4 nkat. L-1 and 1063.8± 10.0 nkat. L-1 . Statistically,the proteins of group Ⅱ and group Ⅲ were significantlyhigher than those of control group (P＜0.05), whereas theprotein production of group Ⅰ, AIb, ALT and LDH were 5.3±0.2 g. L-1, 205.4±3.3 nkat. L-1 and 1035.4± 116.9 nkat. L-1,were similar to those of control group. At the same time,TBE and MTT analysis indicated that there was no significantcell viability difference between electroporation groups andcontrol group.CONCLUSION: This single-pulse electroporation procedureperformed at Iow voltage (220-400 V) but with highcapacitance (950 μF) is one of the optimal choices tointroduce foreign genes into primary rat hepatocyte.展开更多
文摘胰腺导管腺癌(pancreatic ductal adenocarcinoma,PDAC)是恶性程度最高的肿瘤之一。越来越多的证据表明包括立体定向放射治疗(stereotactic body radiotherapy,STRT)和射频消融在内的局部治疗可能成为不可手术切除的PDAC患者的新选择。由于PDAC的单独免疫治疗效果欠佳,近年来越来越多的研究发现局部治疗可以改变肿瘤局部及远处转移灶的免疫微环境,并增加免疫治疗敏感性。可以预期,局部治疗联合免疫治疗可能提高疗效。本文主要介绍PDAC局部治疗联合免疫治疗的机制及相关临床研究进展。
基金the National Natural Science Foundation of China,No.39670340
文摘AIM: To investigate the effects of electroporation on primaryrat hepatocyte and to optimize the electroporation conditionsintroducing foreign genes into primary hepatocytes.METHODS: A single-pulse procedure was performed at Iowvoltage (220-400 V) but with high capacitance (500-950 μF).Hepatocytes were divided into 4 groups according to theelectroporation conditions: group Ⅰ, 220 V and 500 μF;group Ⅱ, 220 Vand 950 μF; group Ⅲ, 400 V and 950 μF,and group Ⅳ.The control group was freshly isolatedhepatocytes and directly cultured under the same conditionsas those of electroporation groups. The effects ofelectroporation on primary rat hepatocytes were detectedby trypan blue exclusion (TBE) and MTT analysis. Besides,albumin (AIb), alanine transaminase (ALT) and lactatedehydrogenase (LDH) in the supernatants of culturedhepatocytes were measured by biochemical assay.RESULTS: Between day 1 and day 15 after incubation,primary rat hepatocytes of each electroporation groupappeared normal, being the same with those of controlgroup. TBE staining showed that slight hepatocyte damageand high survival rate were found in the electroporationgroups and the control group. Cultured for 3, 7, 11 and 15days, hepatocyte viability was approximatly 92.6±2.5 %,89.5±3.3 %, 82.0±3.5 % and 74.3±1.2 %, respectively.MTT analysis indicated that the viabilities of hepatocyteshad no significant difference between each electroporationgroup, and those were similar to that of control group. Atthe 36th hour after electroporation, AIb, ALT and LDH in thesupernatants of control group were 5.3±0.1 g. L-1, 183.7±8.4 nkat. L-1 and 896.8±58.5 nkat. L-1; those of group Ⅱwere 5.7±0.1 g. L-1, 215.4±16.7 nkat. L-1 and 1063.8±51.8 nkat. L-1; and those of group Ⅲ were 5.80.2 g. L-1,217.1 ± 8.4 nkat. L-1 and 1063.8± 10.0 nkat. L-1 . Statistically,the proteins of group Ⅱ and group Ⅲ were significantlyhigher than those of control group (P＜0.05), whereas theprotein production of group Ⅰ, AIb, ALT and LDH were 5.3±0.2 g. L-1, 205.4±3.3 nkat. L-1 and 1035.4± 116.9 nkat. L-1,were similar to those of control group. At the same time,TBE and MTT analysis indicated that there was no significantcell viability difference between electroporation groups andcontrol group.CONCLUSION: This single-pulse electroporation procedureperformed at Iow voltage (220-400 V) but with highcapacitance (950 μF) is one of the optimal choices tointroduce foreign genes into primary rat hepatocyte.
