Autologous CD34^+ and CD133^+ stem cells transplantation in patients with end stage liver disease

AIM:To assess the utility of an autologous CD34 + and CD133 + stem cells infusion as a possible therapeutic modality in patients with end-stage liver diseases.METHODS:One hundred and forty patients with endstage liver diseases were randomized into two groups.Group 1,comprising 90 patients,received granulocyte colony stimulating factor for five days followed by autologous CD34 + and CD133 + stem cell infusion in the portal vein.Group 2,comprising 50 patients,received regular liver treatment only and served as a control group.RESULTS:Near normalization of liver enzymes and improvement in synthetic function were observed in 54.5% of the group 1 patients;13.6% of the patients showed stable states in the infused group.None of the patients in the control group showed improvement.No adverse effects were noted.CONCLUSION:Our data showed that a CD34 + and CD133 + stem cells infusion can be used as supportive treatment for end-stage liver disease with satisfactory tolerability.

Circulating tumor and cancer stem cells in hepatitis C virusassociated liver disease

AIM:To assess the role of circulating tumor cells(CTCs)and cancer stem cells(CSCs)in hepatitis C virus(HCV)-associated liver disease.METHODS:Blood and/or tissue samples were obtained from HCV(genotype 4)-associated hepatocellularcarcinoma patients(HCC;n=120),chronic hepatitis C patients(CH;n=30)and 33 normal control subjects(n=33).Serum levels of alpha-fetoprotein(AFP),alkaline phosphatase,and alanine and aspartate aminotransferases were measured.Cytokeratin 19(CK19)monoclonal antibody was used to enumerate CTCs,and CD133 and CD90 were used to enumerate CSCs by flow cytometry.The expression levels of the CSCs markers(CD133 and CD90)as well as telomerase,melanoma antigen encoding gene 1(MAGE1)and MAGE3 were assessed by RTPCR and quantitative real-time polymerase chain reactions.The number of CTCs and/or the expression levels of CK19,CD133,telomerase,MAGE1 and MAGE3 were correlated to the standard clinicopathologic prognostic factors and disease progression.RESULTS:Levels of AFP,alkaline phosphatase and aspartate aminotransferase were significantly different among the HCC,CH and control groups(P<0.001),whereas alanine aminotransferase differed significantly between patient(HCC and CH)and control groups(P<0.001).At the specified cutoff values determine by flow cytometry,CK19(49.8),CD90(400)and CD133(73)were significantly higher in the blood of HCC patients compared to those in the CH and control groups(P<0.001).On the other hand,CD133 at a 69.5 cutoff was significantly higher in the CH compared to the control group(P≤0.001).Telomerase,MAGE1 and MAGE3RNA were expressed in 55.71%,60.00%and 62.86%of the HCC patients,respectively,but were not detected in patients in the CH or control groups,which were statistically significant(Ps<0.001).The expression levels of telomerase,CD90,MAGE3,CD133 and CK19 were all significantly associated with high tumor grade and advanced stage in HCC patients(all P s<0.05).CONCLUSION:CTC counts and AFP,CK19,telomerase,and MAGE1/MAGE3 expression predict disease progression in patients with HCV,whereas telomerase,MAGE3,CD90,CD133 and CK19 are prognostic mark-

Role of relevant immune-modulators and cytokines in hepatocellular carcinoma and premalignant hepatic lesions

AIM To assess the levels of different immune modulators in patients with hepatocellular carcinoma(HCC),in relation to other hepatic diseases.METHODS Eighty-eight patients were included in the current study and represented patients with HCC(20),liver cirrhosis(28) and chronic hepatitis(CH;25),and normal controls(NC;15).Peripheral blood was isolated for immunophenotyping of active myeloid dendritic cells(m DCs;CD1 c and CD40),mature inactive myeloid cells(CD1 c and HLA),active plasmacytoid cells(p DCs;CD303 and CD40),mature inactive p DCs(CD30 and HLA),active natural killer(NK) cells(CD56 and CD161),active NK cells(CD56 and CD314) and inactive NK cells(CD56 and CD158) was done by flow cytometry.Serum levels of interleukin(IL)-2,IL-10,IL-12,IL-1β,interferon(IFN)-α,IFN-γ and tumor necrosis factor(TNF)-αR2 were assessed by ELISA.RESULTS Active m DCs(CD1 C+/CD40+) and inactive m DCs(CD1 c+/HLA+) were significantly decreased in HCC patients in relation to NC(P < 0.001).CD40+ expression on active p DCs was decreased in HCC patients(P < 0.001),and its level was not significantly changed among other groups.Inactive p DCs(CD303+/HLA+),inactive NKs(CD56+/CD158+) and active NKs(CD56+/CD161+) were not statistically changed among the four groups studied;however,the latter was increased in CH(P < 0.05).NKG2 D was statistically decreased in HCC,CH and cirrhosis(P < 0.001),and it was not expressed in 63%(12/20) of HCC patients.There was significant decrease of IL-2,IFN-α and IFN-γ(P < 0.001),and a significant increase in IL-10,IL-1β,and TNF-αR2(P <0.01,P < 0.001 and P < 0.001;respectively) in HCC patients.There was inverted correlation between IL-12 and IL-1β in HCC(r =-0.565,P < 0.01),with a strong correlation between p DCs(CD303+/CD40+) and NKs(CD56+/CD161+;r = 0.512,P < 0.05) as well as inactive m DCs(CD1 c+/HLA+) and inactive NK cells(CD56+/CD158+;r = 0.945,P < 0.001).CONCLUSION NKG2 D,CD40,IL-2 and IL-10 are important modulators in the development and progression of HCC.

Early detection of hepatocellular carcinoma co-occurring with hepatitis C virus infection: A mathematical model

AIM: To develop a mathematical model for the early detection of hepatocellular carcinoma(HCC) with a panel of serum proteins in combination with α-fetoprotein(AFP).METHODS: Serum levels of interleukin(IL)-8, soluble intercellular adhesion molecule-1(s ICAM-1), soluble tumor necrosis factor receptor Ⅱ(s TNF-RⅡ), proteasome, and β-catenin were measured in 479 subjects categorized into four groups:(1) HCC concurrent with hepatitis C virus(HCV) infection(n = 192);(2) HCV related liver cirrhosis(LC)(n = 96);(3) Chronic hepatitis C(CHC)(n = 96); and(4) Healthy controls(n = 95). The R package and different modules for binary and multi-class classifiers based on generalized linear models were used to model the data. Predictive power was used to evaluate the performance of the model. Receiver operating characteristic curve analysis over pairs of groups was used to identify the best cutoffs differentiating the different groups. RESULTS: We revealed mathematical models, based on a binary classifier, made up of a unique panel of serum proteins that improved the individual performance of AFP in discriminating HCC patients from patients with chronic liver disease either with or without cirrhosis. We discriminated the HCC group from the cirrhotic liver group using a mathematical model(-11.3 + 7.38 × Prot + 0.00108 × s ICAM + 0.2574 ×β-catenin + 0.01597 × AFP) with a cutoff of 0.6552, which achieved 98.8% specificity and 89.1% sensitivity. For the discrimination of the HCC group from the CHC group, we used a mathematical model [-10.40 + 1.416 × proteasome + 0.002024 × IL + 0.004096 × s ICAM-1 +(4.251 × 10-4) × s TNF + 0.02567 ×β-catenin + 0.02442 × AFP] with a cutoff 0.744 and achieved 96.8% specificity and 89.7% sensitivity. Additionally, we derived an algorithm, based on a binary classifier, for resolving the multi-class classification problem by using three successive mathematical model predictions of liver disease status. CONCLUSION: Our proposed mathematical model may be a useful method for the early detection of different statuses of liver disease co-occurring with HCV infection.