Objective To evaluate the effect of the combination of lamivudine and adefovir dipivoxil on the treatment of decompensated hepatitis B cirrhosis patients. Methods One hundred thirty-six patients with decompensated hepatitis B cirrhosis were randomly administered the combination of lamivudine and adefovir dipivoxil (combination group), only lamivudine( LAM group)and only adefovir dipivoxil (ADV group). The liver founction tests, ChildPugh scores and the rate of negative conversion for the serum HBV-DNA were measured at baseline,six months after therapy and 12 months after therapy. Results The rate of negative conversion for the serum HBV-DNA ( ＜ 1 ×103cop/ml)of combination group were 78.3% and 85.2% respectively at six and twelve months after therapy. The rates of negative conversion for the serum HBV-DNA of LAM group were 62. 1% and 57.8% respectively at six and twelve months after therapy. The rates of negative conversion for the serum HBV-DNA of ADV group were 41.3% and 53.6% respectively at six and twelve months after therapy. Compared with the LAM and ADV group, The rates of negative conversion for the serum HBV-DNA of combination group were higher. In addition, the rates of better conversion of liver function tests and Child-Pugh scores were higher in combination group than LAM and ADV groups.Conclusion The combination of lamivudine and adefovir dipivoxil could more effectively improve liver founction tests, Child-Pugh scores and the rates of negative conversion for the serum HBV-DNA in patients with decompensated hepatitis B cirrhosis than only lamivudine and only adefovir dipivoxil.

Objective To investigate the effect of dual-specificity phosphatase-2 (DUSP2) on the cell proliferation and apoptosis in the gastric cancer and its mechanisms. Methods Firstly, the effects of different expressions of DUSP2 on the overall survival of 876 gastric cancer patients were analyzed by online analysis tool KM plotter, and the expressions of DUSP2 in various gastric cancer cell lines (MKN-45, SGC-7901, HGC-27 and N-87) were verified. Secondly, DUSP2 overexpressed lentiviral vector was constructed, and MKN-45 was transfected by packaged virus. DUSP2-overexpression gastric cancer cell line was gained by drug screening. Meanwhile, gastric cancer cells infected with empty vector virus were used as control. Then the effect of DUSP2 upregulation on the proliferation ability of gastric cancer cells was evaluated by MTS cell proliferation assay, and the apoptosis was determined by Annexin V-FITC / PI double staining. The protein expressions of DUSP2, ERK, p-ERK (Thr202/Tyr204), P38 and p-P38 were tested by the Western blot analysis. Results Gastric cancer patients with high DUSP2 expression showed a significant survival advantage compared with those with low DUSP2 expression, and DUSP2 levels were decreased in several gastric cancer cell lines. The Western blot analysis revealed that the expression of DUSP2 markedly increased in overexpressed DUSP2 group (experimental group) compared with that of control group. The MTS experiment showed that the cell viability was significantly decreased in experimental group than that of the control group. Correspondingly, the cell apoptosis test showed that the cell apoptosis rate was obviously higher in the experimental group than that of the control group. The results of Western blot assay indicated that p-ERK (Thr202/Tyr204) and p-38 were significantly down-regulated in the experimental group compared with those of control group. Conclusion The over-expression of DUSP2 can efficiently inhibit cell proliferation and promote its apoptosis in gastric cancer cells, and the mechanism is related to DUSP2 inhibiting the phosphorylation levels of ERK and P38.

OBJECTIVE: To investigate the regulatory effects of miR-26a-5p on the osteogenic differentiation of adipose-derived mesenchymal stem cells (ADSCs) by regulating cAMP response element binding protein 1 (CREB1). METHODS: The adipose tissues of four 3-4 weeks old female C57BL/6 mice were collected and the cells were isolated and cultured by digestion separation method. After morphological observation and identification by flow cytometry, the 3rd-generation cells were subjected to osteogenic differentiation induction. At 0, 3, 7, and 14 days after osteogenic differentiation induction, the calcium deposition was observed by alizarin red staining, ALP activity was detected, miR- 26a-5p and CREB1 mRNA expressions were examined by real-time fluorescence quantitative PCR, and CREB1 protein and its phosphorylation (phospho-CREB1, p-CREB1) level were measured by Western blot. After the binding sites between miR-26a-5p and CREB1 was predicted by the starBase database, HEK-293T cells were used to conduct a dual-luciferase reporter gene experiment to verify the targeting relationship (represented as luciferase activity after 48 hours of culture). Finally, miR-26a-p inhibitor (experimental group) and the corresponding negative control (control group) were transfected into ADSCs. Alizarin red staining, ALP activity, real-time fluorescent quantitative PCR (miR-26a-5p) and Western blot [CREB1, p-CREB1, Runt-related transcription factor 2 (RUNX2), and osteocalcin (OCN)] were performed at 7 and 14 days after osteogenic induction culture. RESULTS: The cultured cells were identified as ADSCs. With the prolongation of osteogenic induction culture, the number of calcified nodules and ALP activity significantly increased ( P<0.05). The relative expression of miR-26a-5p in the cells gradually decreased, while the relative expressions of CREB1 mRNA and protein, as well as the relative expression of p-CREB1 protein were increased. The differences were significant between 7, 14 days and 0 day ( P<0.05). There was no significant difference in p-CREB1/CREB1 between different time points ( P>0.05). The starBase database predicted that miR-26a-5p and CREB1 had targeted binding sequences, and the dual-luciferase reporter gene experiment revealed that overexpression of miR-26a-5p significantly suppressed CREB1 wild-type luciferase activity ( P<0.05). After 7 and 14 days of osteogenic induction, compared with the control group, the number of calcified nodules, ALP activity, and relative expressions of CREB1, p-CREB1, OCN, and RUNX2 proteins in the experimental group significantly increased ( P<0.05). There was no significant difference in p-CREB1/CREB1 between the two groups ( P>0.05). CONCLUSION Knocking down miR-26a-5p promoted the osteogenic differentiation of ADSCs by up-regulating CREB1 and its phosphorylation.
Animals ; Female ; Mice ; Cell Differentiation ; Cells, Cultured ; Core Binding Factor Alpha 1 Subunit/metabolism* ; Cyclic AMP Response Element-Binding Protein/metabolism* ; Mesenchymal Stem Cells ; Mice, Inbred C57BL ; MicroRNAs/metabolism* ; Osteocalcin/metabolism* ; Osteogenesis/genetics* ; RNA, Messenger/genetics*