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Objective To employ high-throughput next generation sequencing (NGS) for analyzing the expression of lneRNAs and mRNAs in donor samples from pediatric living donor liver transplantation and search differentially expressed lncRNAs and drag metabolic gene for individualized guidance of immunosuppressive agents.Methods Between October 2016 and December 2017,10 liver tissue specimens from living donor liver transplantation children were collected and divided into fast and slow metabolic groups (n =5 each) according to the postoperative profiles of drug metabolism.Samples were assayed for high-throughput NGS.Target analysis was used for functional pathways and screening target genes prediction.Results There were differentially expressed 908 mRNAs and 1228 lncRNAs between slow metabolic and fast metabolic groups (P＜0.05).According to the abundance and difference,22 up-regulated and 18 down-regulated mRNAs,13 up-regulated and 24 down-regulated lncRNAs were selected.In addition to CYP3A5,CYP2C19,CYP1A2 and UGT1A1 might affect the metabolism of tacrolimus.At the same time,NONHSAT108617.2 in differemially expressed lncRNAs might regulate the expression of CYP3A5 gene.Conclusions This study has comprehensively analyzed the expression of lncRNAs in donor liver from pediatric liver transplantation.Some differentially expressed drug metabolism related genes may affect tacrolimus metabolism in vivo and thus the postoperative use of immunosuppressive drugs.
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Objective To observe the efficacy of different degrees of portal vein ligation on portal pressure and liver regeneration of the unligated lobe in rats.Methods Seventy-two healthy male SpragueDawley rats were randomly divided into three groups:group A (sham,n =24);group B (n =24) undergoing 70％ portal vein ligation;group C (n =24) undergoing 90％ portal vein ligation.And then the portal pressure and liver regeneration rate (HRR) of the unligated lobe were detected immcdiately and postoperatively at each observation time point in each group.The serum alanine aminotransferase (ALT),aspartate aminotransferase (AST),hepatic proliferating cell nuclear antigen (PCNA) were compared at each observation time point,and the histological changes were observed by HE staining.Results The HRR of the unligated lobe in group B and group C increased obviously postoperatively at each time point,and the HRR in group C was significantly higher than that in group B [(220.1 ± 4.3) ％,(246.3 ± 5.6) ％,(261.4 ±2.3)％ vs (128.2 ±3.7)％,(143.4 ±8.7)％,(150.7 ±7.0)％,P＜0.05].The serum ALT and AST increased obviously on day 1 and then gradually declined,and the serum ALT and AST in group C was significantly higher than those in group B on day 1 [(821.7 ± 158.3) U/L,(1 372.0 ± 376.2) U/L vs (398.6 ± 80.4) U/L,(860.4 ± 80.0) U/L,P ＜ 0.05].The immediate portal pressure in both groups were obviously increased postoperatively and then gradually declined,and the portal pressure in group C was higher than that in group B at each observation time point [(23.5 ± 1.1)cmH2O,(18.8 ±0.9)cmH2O,(17.8±1.0)cmH2O,(16.6 ±1.0)cmH2O,(15.9±1.3)cmH2O vs (17.4 ±1.0)cmH2O,(16.5 ±1.2)cmH2O,(15.3±1.0)cmH2O,(10.2±1.2)cmH2O,(10.0±1.1)cmH2O,P＜0.05].ThePCNA index in group C was higher than that in group B on day 1 and3 [(21.5 ±1.1)％,(28.2±1.3)％ vs (12.8 ± 2.1) ％,(18.8 ± 1.9) ％,P ＜ 0.05].More foca1 necrosis of the unligated lobe were observed in group C on day 1,which were more than those in group B.Conclusion Higher degree of portal vein ligation could cause higher portal pressure,which leads to the greater regeneration of the unligated lobe.
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Objective To investigate the effect of portal vein ligation combined with in situ splitting on liver regeneration in rats .Methods Seventy-five healthy male Sprague-Dawley rats were selected and randomly assigned into sham operation group ( S) , portal vein ligation group ( PVL) and portal vein ligation combined with in situ splitting group ( ALPPS) .On 1 d, 3 d, 7 d, 10 d, 14 d after operation , the hepatic regeneration rate ( HRR) of right median lobe was calculated , the serum alanine aminotransferase ( ALT) , aspartate aminotransferase (AST), IL-6, HGF, VEGF were detected.mRNA of IL-6, HGF, TNF-α, TGF-βwas assayed by real-time PCR, and the hepatic proliferating cell nuclear antigen ( PCNA) labeling index was evaluated by immunohistochemistry .Results Comparing with PVL group , the HRR of the right median lobe obviously increased on day 3, 7, 10 and 14 in ALPPS group (P<0.05), and ALT and AST level were increased on 1 d (P<0.05).On day 1 and 3, the content of serum IL-6, HGF and VEGF were all in-creased in ALPPS group [(70.7 ±14.6) pg/ml vs.(134.2 ±31.4) pg/ml; (0.70 ±0.04) ng/ml vs. (0.74 ±0.02) ng/ml;(82.1 ±12.6) pg/ml vs.(103.5 ±14.7) pg/ml], respectively (P<0.05).The mRNA expression of IL-6, HGF, TNF-α, TGF-βand the PCNA labeling index were also increased in ALPPS group in comparison with those in PVL group on day 1 and 3 (P<0.05).All the indexes in the two groups were all higher than those in the group S ( P<0 .05 ) .Conclusions Portal vein ligation combined with in situ splitting could significantly enhance liver regeneration .The possible mechanisms were related to the inflammation reaction and stress response caused by in situ splitting and up-regulation of cytokines in the regenerating lobe after portal vein ligation combined with in situ splitting , especially IL-6, HGF and TNF-α.
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Objective To establish a rat model of associating liver partition with portal vein ligation for staged hepatectomy (ALPPS) and evaluate the liver function after surgery.Methods Fifty male SD rats were randomly divided into two groups: experimental group (ALPPS group) and control group (PVL group).Selective portal vein ligation in PVL group was performed on the caudal lobe, left lateral and left median lobes, while the right lobe, the right median lobe was preserved to regenerate.ALLPS group was treated in the same way as PVL group, but also underwent liver partition in situ.After surgery, 5 rats were sacrificed on day 1, 3,7, 10 and 14 in each group to observe the weight of body and the right median lobe,respectively.The venous blood and liver tissue were obtained for testing alanine aminotransferase (ALT),aspartate aminotransferase (AST), serum albumin (ALB), total bilirubin (TBil) and observing the histological changes in liver injury after surgery.Results After surgery, the body weight decreased progressively, but then increased in both groups.Since the first day after surgery, the body weight began to decrease,reached the lowest value on 3 d, and then on day 7 the body weight in PVL group returned to preoperative levels.However, the body weight was still lower than that before surgery [(3.7 ± 2.7) ％ vs (-3.0 ± 1.9)％, P＜0.05].On day 3, 7, 10 and 14, the hepatic regeneration rate (HRR) of the fight median lobe in ALPPS group was obviously higher than that in PVL group (P ＜ 0.05).On day 1, the serum ALT and AST levels in two groups were elevated dramatically and then gradually decreased, which in ALPPS group were significantly higher than those in PVL group (P ＜ 0.05).There were no significant differences at other time points.On day 1 and 3, the serum ALB in ALPPS group was obviously lower than that in PVL group [(25.4±1.7)g/Lvs (31.4±1.5)g/L, P＜0.05;(25.0±2.0)g/Lvs (31.8±1.5)g/L, P＜ 0.05], respectively.More focal necrosis of liver were observed in ALPPS group on day 1, which were more than those in PVL group.Conclusions This method could successfully establish a ALPPS rat model and proved that ALPPS could induce accelerated hepatic regeneration and more severe early hepatocyte injury compared with PVL.This ALPPS experimental model provides a basis for further research on ALPPS, especially for clarifying the mechanisms of liver regeneration and tumor recurrence, and exploring the reasons for various ALPPS related complications, which play a significant role in its clinical application.