Genetic testing and analysis of 2 cases of trisomy 11 mosaicism / 南方医科大学学报

Trisomy 11 mosaicism is clinically rare, for which making diagnostic and treatment decisions can be challenging. In this study, we used noninvasive prenatal testing, chromosome karyotype analysis, chromosome microarray analysis, copy number variation sequencing and fluorescence in situ hybridization for detecting trisomy 11 mosaicism in two cases and provided them with genetic counseling. In one of the cases, the fetus with confined placental mosaicism trisomy 11 presented with severe growth restriction and a placental mosaic level of 44%, and pregnancy was terminated at 25+3 weeks of gestation. In the other case with true low-level fetal mosaicism of trisomy 11, the pregnancy continued after exclusion of the possibility of uniparental disomy and structural abnormalities and careful prenatal counseling. The newborn was followed up for more than one year, and no abnormality was found. Noninvasive prenatal testing is capable of detecting chromosomal mosaicism but may cause missed diagnosis of true fetal mosaicism. For cases with positive noninvasive prenatal testing but a normal karyotype of the fetus, care should be taken in prenatal counseling and pregnancy management.

Mechanism and progress of the research on the treatment of bacterial infection with bacteriophage lysin / 基础医学与临床

Bacteriophage lysin is characterized by high stability, wide bactericidal activity and efficacy, and safty. It is able to lyse bacteria specifically and is not susceptible to bacterial resistance. In the presence of phage, phage infecting bacteria and coding for endolysin then it lyse the bacteria, In the absence of the phage, holin assists the endolysin lysis the bacteria from external. In addition, the research progress on the treatment of Gram-positive bacteria, such as methicillin-resistant Staphylococcus aureus, Streptococcus pyogenes, and Gram-negative bacteria, such as Acinetobacter baumannii and Pseudomonas aeruginosa, was also discussed in this paper.

Expression and bioinformatic analysis of miR-29 family, target gene IFN-γ in CD4(+) T cells from subjects with latent tuberculosis infection / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of latent and active pulmonary tuberculosis(TB) on expression of miR-29 family and target gene IFN-γ in CD4(+)T cells.</p><p><b>METHODS</b>Subjects from two hospitals of Weifang were enrolled from March 2012 to December 2012 and divided into three groups: active TB group(30 cases), latent tuberculosis infection(LTBI) group(25 cases) and healthy control group(30 cases). CD4(+) T cells in blood were collected from the three groups.Levels of miR-29a, miR-29b and miR-29c were measured by nucleic acid hybridization and RT-qPCR.Expression of IFN-γ was analyzed by RT-qPCR. Target genes of miR-29 family were predicted with both TargetScan and PicTar.GO annotation and pathway overrepresentation were further analyzed with David database and Cytoscape.</p><p><b>RESULTS</b>Levels of miR-29a, miR-29b and miR-29c showed significant differences among the three groups(P < 0.05): levels of miR-29b and miR-29c in the active TB group(561.63 ± 65.36, 281.85 ± 42.78) were higher than the healthy controls(260.74 ± 38.69, 128.21 ± 19.98), but lower than the LTBI group(2030.29 ± 321.68, 620.93 ± 79.14); expression of miR-29a in the healthy control group(913.95 ± 104.73) were higher than the active TB group(323.37 ± 54.38), but lower than the LTBI group(4782.13 ± 567.81).Level of IFN-γ showed significant differences among the three groups(P < 0.05): level of IFN-γ in the LTBI group(0.45 ± 0.09) were lower than the healthy controls(1.00), but higher than the active TB group(0.11 ± 0.03). The target genes of miR-29 family mainly existed in molecular function such as extracellular matrix structural constituent and transcription regulator activity.In KEGG pathway, the gene set mostly existed in signaling pathway such as Focal adhesion,ECM-receptor interaction and mTOR signaling pathway.</p><p><b>CONCLUSION</b>The expression of miR-29 family was increased and target gene IFN-γ in CD4(+) T cells was decreased by latent and active pulmonary TB, which might play important role in alteration of signal pathway.</p>

Effect of human granulysin on apoptosis, mitochondrial transmembrane potential and cytochrome C release of SMMC-7721 cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To construct a plasmid carrying granulysin (GLS) and to study the effect of the GLS on apoptosis, mitochondrial transmembrane potential and cytochrome C release of SMMC-7721 cells.</p><p><b>METHODS</b>The coding sequence of the GLS was amplified from the total RNA of human CTL cells, and it was inserted into pBudCE4.1 plasmid and then it was used to transfect SMMC-7721 cells. The expression of GLS was detected by RT-PCR and confirmed by immunocytochemistry method. Cell apoptosis was ascertained by Hoechst staining and electron microscopy; mitochondrial transmembrane potential was detected using Mitocapture and cytochrome C release was studied using Western blot.</p><p><b>RESULTS</b>Recombinant pBudCE4.1/GLS plasmid was successfully constructed. GLS protein was successfully expressed in the SMMC-7721 cells and it induced apoptosis of the SMMC-7721 cells, and at the same time, mitochondrial transmembrane potential was reduced and cytochrome C was released from mitochondria into the cytosol.</p><p><b>CONCLUSIONS</b>GLS gene carried by recombinant plasmid could express in SMMC-7721 cells and induce cells apoptosis. The change of mitochondrial transmembrane potential and the release of cytochrome C might be one of the key factors of apoptosis induced by GLS.</p>

A quantitative analysis of mitochondrial protein differential expressions in hydroxycamptothecin-treated hepatoma cells / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the differentially expressed mitochondrial proteins in hydroxycamptothecin (HCPT)-treated SMMC-7721 cells by using quantitative proteome.</p><p><b>METHODS</b>SMMC-7721 cell apoptosis was induced by HCPT and the mitochondria were isolated with a mitochondria isolation kit. Mitochondrial proteins labeled with a cleavable isotope-coded affinity tag were identified and quantified using two-dimensional liquid chromatography/tandem mass spectrometry.</p><p><b>RESULTS</b>Highly purified mitochondria were obtained. Seventy-four mitochondrial proteins, which were statistically significantly altered (P less than 0.05) in HCPT-treated cells, were identified and analyzed. A total of 42 proteins were significantly down-regulated, and 32 were up-regulated in the cells that responded to apoptosis. The functions of these proteins were likely involved in cell energy metabolism, nucleic acid translation and transcription, cytoskeleton, etc.</p><p><b>CONCLUSION</b>Our results about the information of differentially expressed mitochondrial proteins in HCPT-treated cells and the control cells will help to understand the mechanism by which HCPT induces cell apoptosis. The integrated techniques we used in this study will be helpful to the investigation of subcellular quantitative proteomics.</p>

Proteomic analysis of mitochondrial proteins in hydroxycamptothecin-treated SMMC-7721 cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the differentially expressed mitochondrial proteins in hydroxycamptothecin (HCPT)-treated SMMC-7721 cells by comparative proteomic analysis.</p><p><b>METHODS</b>Apoptosis of SMMC-7721 cells were induced by using HCPT and their mitochondria were isolated with a mitochondria isolation kit for cultured cells. Three different solubility protein fractions were extracted with ReadyPrep Sequential Extraction Kit and were separated by two-dimensional gel electrophoresis (2-DE). PDQuest software was used to differentiate mitochondrial proteins between control cells and HCPT-treated cells. Matrix assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS) was used to identify some of the different proteins.</p><p><b>RESULTS</b>Highly purified mitochondria and high resolution 2-DE patterns of the proteins were obtained. Forty-four mitochondrial protein spots from the HCPT-treated cells showed different expressions compared to those of the control cells. Twenty of the different protein spots were analyzed by MALDI-TOF-MS.</p><p><b>CONCLUSION</b>Differently expressed mitochondrial proteins in HCPT-treated cells and control cells were obtained in this study. This will be of help to understand the mechanism by which HCPT induces cell apoptosis.</p>

The transportation of apoptosis-inducing factor and DNA damage during hydroxycamptothecin-induced apoptosis of SMMC-7721 cells / 肿瘤

Objective: To investigate the relationship between transportation of apoptosis-inducing factor (AIF) from mitochondria to nucleus and DNA damage at different time points during apoptosis of SMMC-7721 cells induced by hydroxycamptothecin (HCPT). Methods: Cells were harvested at different time points after HCPT treatment. The mitochondrium, cytosol, and nucleus were isolated with mitochondria isolation kit. These fractions were used for western blot. Transportation of AIF from mitochondrium to the nucleus at different time points was observed by confocal microscopy. At the same time, DNA damage was detected by TUNEL assay. Results: Western blot showed that transportation of AIF from mitochondrium to nucleus was not observed after treatment with HCPT for 1 h. However, AIF immunoreactivity was significantly increased in the nuclear fraction at 6, 12 and 24 h after HCPT treatment. Confocal microscopy demonstrated that AIF was released from mitochondrium at 6 h after HCPT treatment. The massive transportation of AIF to the nucleus occurred at 12 and 24 h after HCPT treatment. At the same time, DNA damage was observed by TUNEL assay. Conclusion: AIF release is one of the early biochemical changes of apoptosis of SMMC-7721 cells after HCPT treatment. The DNA damage occurrs in parallel with AIF translation to nucleus.

Effect of hydroxycamptothecin on apoptosis-inducing factor (AIF) expression and on AIF translocation in human hepatocellular cancer cell SMMC-7721 / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the effect of hydroxycamptothecin (HCPT) on apoptosis-inducing factor (AIF) expression and AIF translocation from mitochondria to the nucleus in human hepatocellular cancer cell SMMC-7721 during apoptosis.</p><p><b>METHODS</b>After treatment with 80 mg/ml of HCPT, the cancer cells were stained with A0/EB to monitor their apoptosis. Their mitochondria was examined with electronmicroscopy and the AIF expression of the cells was tested by RT-PCR and Western blot. The translocation of AIF from mitochondria to the nucleus during apoptosis was analyzed by confocal microscopy.</p><p><b>RESULTS</b>SMMC-7721 cells treated with HCPT showed chromatin condensation, nuclear fragmentation and mitochondria swelling. The mRNA and protein expression of AIF in treated and untreated SMMC-7721 cells were not significantly different. However, cells treated with 80 mg/ml HCPT for 6 h or 12 h showed massive translocation of AIF into the nuclei.</p><p><b>CONCLUSION</b>These results show the important role the mitochondrial pathway of apoptosis plays in HCPT-induced tumor cell death, at least in SMMC-7721 cells.</p>