BACKGROUND:Immunotherapy with autologous immune cel s has been developed as a major adjuvant therapy for malignant tumors, but its mechanism of action has not been elucidated. OBJECTIVE:To investigate the relationship between cytokine-induced kil er cel secretion and apoptosis in human liver cancer stem cel s. METHODS:Human liver cancer stem cel s, HepG2 cel s, were isolated and enriched using serum-free suspension method. The peripheral blood mononuclear cel s from patients with liver cancer were induced byγ-interferon, CD3 monoclonal antibody and recombinant human interleukin-2 to form kil er cel s. Passage 1 liver cancer stem cel s were divided into control group (culture alone) and experimental group (co-culture of cytokines-induced kil er cel s and human liver cancer stem cel s). At 48 hours after culture, apoptosis in human liver cancer stem cel s was detected using flow cytometry, and expression of caspase-3 mRNA and protein was detected using RT-PCR and western blot, respectively. RESULTS AND CONCLUSION:The apoptotic rate in the control group was significantly lower than that in the experimental group (P<0.05). The expressions of caspase-3 at mRNA and protein levels were both higher in the experimental group than the control group (P<0.05). Experimental findings show that cytokines-induced kil er cel s can significantly promote apoptosis in human liver cancer stem cel s, and up-regulate the caspase-3 mRNA and protein expressions dramatical y.

BACKGROUND:Bone marrow mesenchymal stem cells can differentiate into hepatic stem cells in a specific environment, and participate in the repair and reconstruction of liver function.OBJECTIVE:To investigate the therapeutic effect of autologous bone marrow mesenchymal stem cell transplantation on decompensated hepatitis B.METHODS:Eighty-four patients with decompensated hepatitis B were randomly divided into two groups:normal group (n=42) with symptomatic treatment, and oral entecavir, Fuzheng Huayu Capsule; stem cell group (n=42) with the left and right hepatic artery transplantation of autologous bone marrow mesenchymal stem cells (1×106/kg) based on conventional treatments. Degree of liver fibrosis, liver function and score on Model for End-Stage Liver Disease (MELD) system, 1-year survival rate of patients were detected and analyzed with statistics before and 12, 24 weeks after treatment.RESULTS AND CONCLUSION:(1) Liver fibrosis after treatment in two groups:hyaluronic acid, laminin, type Ⅲ collagen and type IV collagen levels after treatment were lower than those before treatment in both two groups (P < 0.05), while these indexes in the stem cell group were lower than those in the normal group at 12 and 24 hours after treatment (P <0.05). (2) Liver function:after treatment, decreased alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels were found in both two groups (P < 0.05), and albumin, cholinesterase, prothrombin activity levels were higher than those before treatment (P < 0.05). The alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels in the stem cell group were lower than those in the normal group at 12 and 24 weeks after treatment, while the cholinesterase level was higher in the stem cell group (P < 0.05). (3) MELD scores:MELD scores were both decreased in the two groups after treatment (P < 0.05), and lower in the stem cell group compared with the normal group at 24 weeks after treatment (P < 0.05). (4) The 1-year survival rate was higher in the stem cell group (69%) than the normal group (50%; P < 0.05). To conclude, the use of autologous bone marrow mesenchymal stem cell transplantation in the treatment of hepatitis B can significantly improve the patients' liver fibrosis and liver function, and improve the 1-year survival rate of patients.

Objective To investigate the effect of stem cells transplantation on immune function,liver function and related indexes in the patients with end-stage liver disease(ESLD).Methods A total of 163 cases of ESLD in Nanyang Municipal Central Hospital of Affiliated Hospital of Zhengzhou University were selected and divided into 2 groups by the randomized single blind method.Eighty-one cases in the control group were given the conventional symptomatic treatment,while 82 cases in the observation group received bone marrow mesenchymal stem cell(BMSC) transplantation based on the control group.The changes of immune function,liver function,alpha fetoprotein(AFP),rate of prothrombin activity(PTA) and plasma total protein(TP) level before and after treatment were observed in the two groups.Results The immune function indexes CD3+,CD4+,CD8+,CD4+/CD8+,TP and PTA levels after treatment in the observation group were significantly higher than those before treatment and in the control group (P＜0.05),while the levels of AFP,alanine aminotransferase (ALT),aspartate aminotransferase (AST) and total bilirubin (TBIL)were lower than those before treatment in the same group and the control group(P＜0.05).There was no statistically significant difference in complication occurrence rate between the two groups(P＞0.05).Conclusion BMSC transplantation for treating ESLD can improve the immune function,improves the liver function and reduces the AFP level.

Objective To observe the effect of Interferon-α2b on HEL cells (human erythroleukemia cell line) growth, apoptosis and JAK2 V617F mutation gene expression. Methods HEL cells were placed in RPMI1640 containing 10% FBS and incubated in a cell incubator. Cells in the logarithmic growth phasem were collected, adjusting the cell density to 1 × 105/mL for experimental research. The interferon concentration in five groups were 0, 5 × 105, 10 × 105, 50 × 105, 100 × 105 U/L, with different incubation time (0, 24, 72, 120 h), respectively. The cell growth status in different groups was observed in the inverted optical microscope; MTT was used to detect the inhibition of interferon on HEL cell proliferation. Cell apoptosis was detected by flow cytometry. Fluorescence quantitative PCR was used to detect the mutation gene of JAK2 V617F expression. Results Inhibition rates of Interferon on the HEL cell proliferation in 5 × 105 U/L, 10 × 105 U/L, 50 × 105 U/L, 100 × 105 U/L groups were 18.57%, 25.10%, 42.10%, 57.00%, respectively. JAK2 V617F/GAPDH by fluorescence quantitative was 1.556, 1.213, 0.870 respectively under the concentration of interferon 100 × 105 U/L for 24, 72, 120 h. Conclusions Interferon-α2b can inhibit HEL cells proliferation and induce HEL cells apoptosis. Increasing concentration of interferon increases HEL cell apoptosis rate. Interferon can inhibit JAK2 V617F expression of HEL cells in a dose-dependent manner.