Objective To review the regulation of liver regeneration factors. Methods The literatures about liver regeneration related regulators in recent years were reviewed. Results With further advancement of researches on regulators of liver regeneration in recent years, there were more therapies for treatment of liver-related diseases. Regulators play important roles in the process of liver regeneration, as one of which, cell growth factor plays an essential role in liver cell proliferation, such as the proper expression of TNF-α and IL-6 promoting liver cell proliferation, HGF, TGF-α, EGF, ALR, FS, and others motivating liver cell proliferation, while TGF-β and IL-1 physically terminating liver cell proliferation. Conclusion By strengthening the studies on liver regeneration regulators, new methods may appear for treating liver-related diseases.
Objective To explore the liver regeneration following partial liver transplantation. MethodsPartial liver transplantation in the rats were established, three experimental groups were: Ⅰ=control, partial liver resection; Ⅱ=orthotopic liver transplantation (OLT); Ⅲ=partial orthotopic liver transplantation (POLT). Liver function test, morphological investigations and liver regeneration were performed in different time after transplantation. The regenerative response of transplanted partial liver graft in rats were evaluated by Flow Cytometry and compared it to liver regeneration following resection.Results The serum concentrations of ALT, BILI increased in one week, but returned gradually to normal level within one month after transplantation. Large numbers of mononuclear cells infiltrating into the portal areas. Hepatocyte necrosis was observed on day 14 after transplantation. On day 30, the parenchyma cell showed a nearly normal appearance, bile duct proliferation was seen in portal areas. In addition, after liver resection and POLT some diploid hepatocytes were found. Dilation of the central veins, adjoining sinusoids and interlobar veins were seen in group Ⅲ. The partial liver graft is capable of regeneration similar to the situation following partial hepatectomy. The peak of liver regeneration was seen on day 1,2,4 following a hepatectomy and POLT and OLT, respectively.Conclusion The transplanted liver shows the same and/or enhanced regeneration compared to controls. There are several possible explanations for the slight delay in achieving the maximal regenerative response in rats undergoing the POLT and OLT. These may include damage that is induced by the operation itself, preservation, and reperfusion injuries. These suggest that this be caused by activation of the immune system and it might be related to the regulation of cytokines and hormone.
Objective To investigate the value of retention rate of indocyanine green at fifteen minutes (ICGR15) during hemihepatectomy for evaluation of residual liver reserve function in patients with primary liver carcinoma. Methods During hemihepatectomy, ICGR15 was tested in 44 patients after the hepatic artery and portal vein of resected side were ligated. Child-Pugh score, Child-Pugh classification, and MELD score before operation were tested. After operation, the liver function condition was estimated. Results The incidence of liver dysfunction was significantly lower in ICGR15lt;10% group (17.9%, 5/28) than that in 10%~15% group (75.0%, 12/16), Plt;0.05. There was no significant difference of Child-Pugh score among normal liver function group, mild insufficiency of liver function group, and severe insufficiency of liver function group （Pgt;0.05）. ICGR15 and MELD score in normal liver function group were statistical lower than those in mild insufficiency of liver function group and severe insufficiency of liver function group (Plt;0.05). ICGR15 was significantly lower in Child-Pugh A group than that in Child-Pugh B group (Plt;0.05). Conclusion Intraoperative residual liver ICGR15 may be more precisely compared with Child-Pugh score in evaluation liver reserve function for the patients with primary liver carcinoma and can help to guide liver resection.
Objective To study the effects of peroxisome proliferators-activated receptor (PPAR) γ on the growth of human hepatocellular carcinoma cells and explore the roles of phosphatase and tensin homologue deleted on chromosome ten (PTEN) and phospho-Akt in this process. Methods SMMC-7721 cells were treated with 15-d-PGJ2 or pioglitazone, which were two kinds of PPARγ ligands, at different concentrations. The viability of SMMC-7721 cells was evaluated by MTT assay. The cell cycle was analyzed by flow cytometry. PTEN mRNA level was determined by RT-PCR. The protein expressions of PTEN and pAkt were measured by Western blot analysis. Results It was demonstrated through MTT assay that both 15-d-PGJ2 and pioglitazone had an inhibitory effect on the growth of SMMC-7721 cells in a time- and dose- dependent manner. According to flow cytometry detection, more cells were arrested in G0/G1 phase. Increased expression of PTEN mRNA was detected in 15-d-PGJ2 or pioglitazone-treated cells through RT-PCR. Increased expression of PTEN protein and decreased expression of pAkt were confirmed by Western blot analysis. Conclusion The ligands of PPARγ could inhibit SMMC-7721 cells proliferation in a time- and dose- dependent manner. The upregulation of PTEN may be involved in the underlying mechanism.
ObjectiveTo evaluate the effect of heparin binding epidermal growth factor-like growth factor (HB-EGF) on liver regeneration after partial orthotopic liver transplantation. MethodsFourty SD rats were used to establish the model of partial orthotopic liver transplantation with ameliorated two-cuff technique. Then all the rats were divided into 2 groups: experiment group and control group. Twenty rats of experiment group were administered 500 μg/kg HBEGF via vena caudalis immediately after operation twice a day, while the same volume of saline was administered to the rats in control group. Five rats in each group were selected randomly and killed at the 6th hour, day 2, 4 and 7 after operation, respectively. The serum levels of albumin (Alb) and alanine aminotransferase (ALT) in the blood sample were detected. Every liver was removed and weighed. The expression of Ki67 was detected by using immunohistochemistry assay. The regeneration activity of hepatocytes was evaluated by flow cytometry. ResultsThe wet weights of liver in experiment group were all significantly higher than that in control group at the 6th hour, day 2 and 4 after transplantation (P＜0.05). The serum levels of ALT were significantly lower in experiment group than those in control group at the 6th hour, day 2， 4, 7 after operation (P＜0.05), while the levels of Alb were significantly higher on day 4 and 7. The proliferating index and Ki-67 labeling index of graft in experiment group were higher than those in control group on day 2 and 4 after transplantation （2 d: P＜0.01; 4 d: P＜0.05）. ConclusionHBEGF could promote the regeneration of rat hepatocytes after partial liver transplantation.
【Abstract】ObjectiveTo investigate whether heme oxygenase-1 can alleviate the ischemiareperfusion injury of the aged donor liver. MethodsThe activity of superoxide dismutase (SOD) and catalase (CAT), and the contents of tocopherol (Vit E), ascorbic acid (Vit C) and malondialdehyde (MDA) were measured in the livers of adult SD rats (n=5) and aged SD rats (n=5). The experimental aged donor group (n=30) received intraperitoneal injection of Hemin 24 hours before operation, the control aged donor group(n=30) received saline. The histologic changes and apoptosis in the donor liver were observed. ResultsThe activity of SOD and the contents of Vit E and Vit C decreased significantly in 5 aged rats（P＜0.05）, but the content of MDA increased（P＜0.05）. Before the harvesting of the grafts, the activity of SOD and the contents of Vit E and Vit C increased significantly in rats pretreated with Hemin （P＜0.05） and the content of MDA decreased（P＜0.05）. The apoptotic cells in the livers pretreated with Hemin also decreased significantly after reperfusion（P＜0.05）. ConclusionThe liver of aged rat presents oxidative stress and peroxidative state. Ischemia-reperfusion injury can be alleviated by the induction of HO-1.
ObjectiveTo develop and modify a rat model of partial liver transplantation with hepatic arterialization and to investigate the postoperative histopathologic changes of the grafts. MethodsModified twocuff technique was applied to establish a rat model of partial liver transplantation, and the donor’s celiac artery was anastomosed with the recipient’s right renal artery. The biopsy of the grafts was performed on the day 1,2,4 and 7 after transplantation. Results85.4% of the recipients had survived for one week.Nuclear mitoses were seen on the postoperative day 2 by microscopy, while on the day 4 they were found occasionally. Two nucleus cells were seen on the day 4 and 7. Conclusion The stability of the model is improved by modifying the technique. Liver acupuncture biopsy is a simple, safe and feasible way to be applied in the research of postoperative problems of liver transplantation. Reconstruction of hepatic artery is possible to enhance the regeneration of liver grafts.
【Abstract】Objective To design the hammerhead ribozyme gene according to the hTR sequence in the gallbladder cancer cell, and build it into the eukaryon expression vector pTriEx-4. Methods According to the hTR cDNA sequence, the authors designed the primers and take the hTR template area gene from the gallbladder cancer cells by RT-PCR.The hammerhead ribozyme gene was synthesize according to the result of sequencing, and combine them with eukaryon expressing vector. Identified the exactitude of recombine vector by digestion.Results The 68 bp sequence extracted from the cell through the RT-PCR had the same template sequence comparing with the hTR cDNA. The recombinant plasmid with the hammerhead ribozyme gene was correct by digestion identification. Conclusion The RT-PCR method can extract the gallbladder cancer cell’s hTR gene. We construct the eukaryon expression vector containing the hammerhead ribozyme gene successfully which is the foundation for gene therapy of gallbladder cancer.
【Abstract】ObjectiveTo introduce a new approach for site-specific conjugation of Fab’ fragments of mouse monoclonal antibody (MoAb) to a bifunctional chelator 6-〔p-(bromoacetamido)benzyl〕-1,4,8,11 -tetraazacyclo-tetradecane-N,N′,N″,N-tetraacetic acid (BAT).MethodsFab’ fragments of B43 MoAb were obtained by a simple twostep method： 1, stable F(ab’)2 was produced by pepsin treatment of B43 MoAb. 2, Fab’ was obtained by reduction of the F(ab’)2 with cysteine. Fab’ fragments were directly conjugated with BAT. ResultsFab’ contained about 1.8 free thiol per molecule. The conjugation rate was 74% and each Fab’ fragment carried 1.28 BAT. The F(ab’)2, Fab’ and Fab’-BAT all maintained good biologic activity. ConclusionThis study may provide a simple and efficient method conjugating of MoAb Fab’ fragments to BAT.
Objective To summarize the changes of serum ceruloplasmin levels and urinary copper excretion in Wilson’s disease （WD） after living-related liver transplantation(LRLT) and orthotopic liver transplantation(OLT). Methods From September 2000 to November 2003, 140 cases liver transplantation were performed in our Liver Transplantation Center, LRLT and OLT were carried out in 26 patients with WD, three of them had fulminant hepatic failure and the others had end-stage hepatic insufficiency. All the recipients had low serum ceruloplasmin levels 〔(124.8±22.8) mg/L〕 and high urinary copper excretion 〔(1 524.8±328.6) μg/24h〕 before transplantation. The serum ceruloplasmin levels and urinary copper excretion were within normal limits in 22 donors 〔(230.4±29.6) mg/L〕 and ＜50 μg/24h〕. Results All recipients recovered satisfactorily. After operation 1, 3, 6,12 months, in OLT group, serum ceruloplasmin level and urinary copper excretion were (320.2±36.8) mg/L, (380.4±45.6) mg/L, (360.5±37.6) mg/L, (356.2±27.6) mg/L and (240.4±22.8) μg/24h, (86.5±10.6) μg/24h, (54.2±6.8) μg/24h, (46.8±3.4) μg/24h; While in LRLT group, serum ceruloplasmin levels and urinary copper excretion were (216.8±20.4) mg/L, (248.5±32.6) mg/L, (285.4±44.3) mg/L, (260.2±36.6) mg/L and (380.8±37.6) μg/24h, (150.6±24.5) μg/24h, (75.5±9.6) μg/24h, (60.3±5.8) μg/24h. Conclusion OLT and LRLT are curative procedure in WD manifested as fulminant hepatic failure and/or end-stage hepatic insufficiency. After liver transplantation, the serum ceruloplasmin level can increase to its normal range while urinary copper excretion decreases.