ABSTRACT
Neural stem cells (NSCs) proliferation can be influenced by repetitive transcranial magnetic stimulation (rTMS) in vivo via microRNA-106b-25 cluster, but the underlying mechanisms are poorly understood. This study investigated the involvement of microRNA-106b-25 cluster in the proliferation of NSCs after repetitive magnetic stimulation (rMS) in vitro. NSCs were stimulated by rMS (200/400/600/800/1000 pulses per day, with 10 Hz frequency and 50% maximum machine output) over a 3-day period. NSCs proliferation was detected by using ki-67 and EdU staining. Ki-67, p21, p57, cyclinD1, cyclinE, cyclinA, cdk2, cdk4 proteins and miR-106b, miR-93, miR-25 mRNAs were detected by Western blotting and qRT-PCR, respectively. The results showed that rMS could promote NSCs proliferation in a dose-dependent manner. The proportions of ki-67+ and Edu+ cells in 1000 pulses group were 20.65% and 4.00%, respectively, significantly higher than those in control group (9.25%, 2.05%). The expression levels of miR-106b and miR-93 were significantly upregulated in 600-1000 pulses groups compared with control group (P<0.05 or 0.01 for all). The expression levels of p21 protein were decreased significantly in 800/1000 pulses groups, and those of cyclinD1, cyclinA, cyclinE, cdk2 and cdk4 were obviously increased after rMS as compared with control group (P<0.05 or 0.01 for all). In conclusion, our findings suggested that rMS enhances the NSCs proliferation in vitro in a dose-dependent manner and miR-106b/p21/cdks/cyclins pathway was involved in the process.
Subject(s)
Animals , Rats , Animals, Newborn , Biomarkers , Metabolism , Cell Proliferation , Genetics , Cyclin-Dependent Kinase 2 , Genetics , Metabolism , Cyclin-Dependent Kinase 4 , Genetics , Metabolism , Cyclin-Dependent Kinase Inhibitor p21 , Genetics , Metabolism , Cyclin-Dependent Kinase Inhibitor p57 , Genetics , Metabolism , Cyclins , Genetics , Metabolism , Gene Expression Regulation , Hippocampus , Cell Biology , Metabolism , Ki-67 Antigen , Genetics , Metabolism , Magnetic Fields , MicroRNAs , Genetics , Metabolism , Neural Stem Cells , Cell Biology , Metabolism , Primary Cell Culture , Rats, Sprague-Dawley , Signal TransductionABSTRACT
<p><b>OBJECTIVE</b>To investigate the impact of early rapid virological response on the outcomes of hepatitis B associated acute on chronic liver failure during antiviral treatment.</p><p><b>METHODS</b>106 acute on chronic liver failure patients in our hospital from January 2008 to July 2010 were enrolled in present study retrospectively. Besides internal medicine therapy, all patients received lamivudine (100 mg/d) or entecavir (0.5 mg/d) treatment. The profile of liver biochemistry, prothrombin time activity and viral load were detected at baseline and week 4, respectively. The patients were divided into HBV DNA negative group and HBV DNA positive group according to the viral load at week 4. The clinical features and treatment outcomes were compared between groups. Frequency variables were compared by x2 test or Fisher's exact test. Continuous variables were compared using independent samples T-test. The factors that impact on the treatment outcomes were determined using stepwise multivariate logistic regression analysis.</p><p><b>RESULTS</b>At the week 4, the TBil and PTA in HBV DNA positive group [(261.6+/-205.6)mumol/L and 44.7%+/-19.7%, respectively] were significantly different from those in HBV DNA negative group [(160.1+/-173.4) mumol/L and 56.8%+/-23.1%, respectively] ( t = 2.190 and -2.077, respectively, P less than 0.05). The non-effective rate of HBVDNA positive group (50%, 9/18) was significantly higher than that of HBV DNA negative group (14.8%, 13/88) (x2 = 9.235, P less than 0.01). By using stepwise multivariate logistic regression analysis, the disease stage and HBV DNA undetectable at week 4 were the independent factor. The OR values of disease stage and HBV DNA undetectable were 6.559 and 0.209, respectively, and 95% CI was 2.316~18.576 and 0.058~0.747, respectively.</p><p><b>CONCLUSION</b>The rapid suppression of viral load by nucleotide analogue may improve the efficacy of hepatitis B associated acute on chronic liver failure treatment. The early rapid virological response within first 4 weeks may contribute to the prediction of the treatment outcomes.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Antiviral Agents , Therapeutic Uses , DNA, Viral , Blood , End Stage Liver Disease , Drug Therapy , Virology , Hepatitis B , Drug Therapy , Liver Failure, Acute , Drug Therapy , Virology , Prognosis , Retrospective Studies , Treatment Outcome , Viral LoadABSTRACT
By exploring the different components of the lysis buffer and optimize the 2-DE conditions,established the best proteomics technical system for Populus euphratica's heteromorphic leaves,while take the heteromorphic leaves in the same blanche as the test materials to find differences between the protein expressions of the leaves.It showed that the lysis solution which containing 2mmol/L thiourea,7mmol/L urea,2% CHAPS,60mmol/L DTT and 0.2% IPG buffer could dissolve the protein better.Through tandem mass spectrum,the results show that heteromorphic leaves are different in photosynthesis and respiration.This research offered valuable informations for understanding the molecular mechanism during leaves development and elucidating the mechanism of the eco-adaptability of Populus euphratica.