Sodium valproate induces mitochondria-dependent apoptosis in human hepatoblastoma cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Sodium valproate inhibits proliferation in neuroblastoma and glioma cells, and inhibits proliferation and induces apoptosis in hepatoblastoma cells. Information describing the molecular pathways of the antitumor effects of sodium valproate is limited; therefore, we explored the mechanisms of action of sodium valproate in the human hepatoblastoma cell line, HepG2.</p><p><b>METHODS</b>The effects of sodium valproate on the proliferation of HepG2 cells were evaluated by the Walsh-schema transform and colony formation assays. Sodium valproate-induced apoptosis in HepG2 cells was investigated with fluorescence microscopy to detect morphological changes; by flow cytometry to calculate DNA ploidy and apoptotic cell percentages; with Western blotting analyses to determine c-Jun N-terminal kinases (JNK), p-JNK, Bcl-2, Bax, and caspase-3 and -9 protein expression levels; and using JC-1 fluorescence microscopy to detect the membrane potential of mitochondria. Statistical analyses were performed using one-way analysis of variance by SPSS 13.0 software.</p><p><b>RESULTS</b>Our results indicated that sodium valproate treatment inhibited the proliferation of HepG2 cells in a dose-dependent manner. Sodium valproate induced apoptosis in HepG2 cells as it: caused morphologic changes associated with apoptosis, including condensed and fragmented chromatin; increased the percentage of hypodiploid cells in a dose-dependent manner; increased the percentage of annexin V-positive/propidium iodide-negative cells from 9.52% to 74.87%; decreased JNK and increased phosphate-JNK protein expression levels; reduced the membrane potential of mitochondria; decreased the ratio of Bcl-2/Bax; and activated caspases-3 and -9.</p><p><b>CONCLUSION</b>Sodium valproate inhibited the proliferation of HepG2 cells, triggered mitochondria-dependent HepG2 cell apoptosis and activated JNK.</p>

Optimization of urinary sample preparation process for real-time PCR detection of BK virus / 第二军医大学学报

Objective: To develop an effective preparation method to improve the sensitivity and accuracy of real-time PCR in detection of BK virus's (BKV's) load in urine samples. Methods: A total of 24 samples documented as positive probes in primary detection were enrolled in this study. The candidate samples were prepared by 4 different approaches: unprocessed urine, BKV's DNA extracted from urine, 1:10 diluted urine, and 1:100 diluted urine; and then they were subjected to real-time PCR examination to obtain the viral load. The data obtained were analyzed by SPSS 11.0. Results: The four different preparation processes for urinary specimens had significant impact on detection results of real-time PCR. Three samples were negative in the unprocessed urine group and 66.7% of its samples had the lowest viral loads compared with the other three groups. Two samples in the 1:100 diluted urine group were negative and 79.2% of its samples had the highest viral loads, but its median load was similar to that of the 1:10 group. Viral gene was detected in all samples in the DNA extraction group and 1:10 diluted urine group, but the loss of the target gene was more severe in the DNA extraction group. Conclusion: The 1:10 diluted urine is better for real-time PCR detection of BKV's load, as it lose less viral gene and is more efficient, easy to perform and economical.

Antisense oligonucleotides targeting protein kinase C alpha inhibits the proliferation of A549 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of antisense oligonucleotides (ASODN) targeting protein kinase C alpha (PKCalpha) on the proliferation of A549 cells.</p><p><b>METHODS</b>PKCalpha ASODN and random oligonucleotides (RODN) were transfected into A549 cells mediated by polyethyleneimine, and the proliferation and clone formation of A549 cells were detected by CCK-8 and clone formation assay, respectively. The expression of PKCalpha in the transfected cells was analyzed by RT-PCR and Western blotting.</p><p><b>RESULTS</b>Compared with those in the control group, PEI group and PEI-RODN group, the proliferation and clone formation of A549 cells treated with ASODN targeting PKCalpha were significantly inhibited (P<0.05). The expressions of PKCalpha mRNA and protein in PKCalpha ASODN-transfected A549 cells were significantly lower than those in the other 3 groups (P<0.05).</p><p><b>CONCLUSION</b>The PKCalpha ASODN mediated by PEI down-regutates the expression of PKCalpha gene and suppress the proliferation and clone formation of A549 cells.</p>