[IL-32 mRNA Expression of Bone Marrow Stromal Cells and Its Correlation with Cell Apoptosis in Patients with Myelodysplastic Syndrome].

OBJECTIVE: To investigate the IL-32 mRNA expression of bone marrow stromal cells and its correlation with apoptosis of bone marrow mononuclear cells in patients with myelodysplastic syndrome (MDS). METHODS: Bone marrow samples from 26 MDS patients and 10 iron deficiency anemia (IDA, as control) patients were collected, RT-PCR was used to detect the IL-32 mRNA expression of bone marrow stromal cells, and the apoptosis of bone marrow mononuclear cells was detected by flow cytometry with Annexin V-FITC/PI dowble staining. The born marrow lymphocytes and NK cells were detected by means of direct immunofluorescence labeling whole blood hemolysis and flow cytometry. RESULTS: IL-32 mRNA expression of bone marrow stromal cells in the MDS patients was significantly higher than that of control group, the IL-32 mRNA expression of bone marrow stromal cells in patients with RA, RAS and RCMD was significantly higher than that in patients with RAEB. There was no obvious difference between RAEB and the control groups. The apoptosis of bone marrow mononuclear cells in MDS group was significantly higher than that in the control group, the apoptosis of bone marrow mononuclear cells in patients with RA, RAS and RCMD was significantly higher than that in RAEB. There was no significant difference between RAEB group and control group. The IL-32 mRNA expression in bone marrow stromal cells significantly correlated with the apoptosis of bone marrow mononuclear cells in MDS patients. The NK cell number in born marrow of MDS patients and the control group had no significant difference. CONCLUSION: The expression of IL-32 mRNA in bone marrow stromal cells significantly relates with the apoptosis of MDS cells, and the secretion of IL-32 by bone marrow stromal cells may be one of the reasons for the apoptosis of MDS bone marrow cells. It is speculated that the abnormal MDS bone marrow microenvironment is involved in the apoptosis of bone marrow cells.

Genome-wide analysis and expression profiling under heat and drought treatments of HSP70 gene family in soybean (Glycine max L.).

Heat shock proteins (HSPs) perform a fundamental role in protecting plants against abiotic stresses. Previous studies have made great efforts in the functional analysis of individual family members, but there has not yet been an overall analysis or expression profiling of the HSP70 gene family in soybeans (Glycine max L.). In this study, an investigation of the soybean genome revealed 61 putative HSP70 genes, which were evaluated. These genes were classified into eight sub-families, denoted I-VIII, based on a phylogenetic analysis. In each sub-family, the constituent parts of the gene structure and motif were relatively conserved. These GmHSP70 genes were distributed unequally on 17 of the 20 chromosomes. The analysis of the expression profiles showed that 53 of the 61 GmHSP70 genes were differentially expressed across the 14 tissues. However, most of the GmHSP70s were differentially expressed in a tissue-specific expression pattern. Furthermore, the expression of some of the duplicate genes was partially redundant, while others showed functional diversity. The quantitative real-time PCR (qRT-PCR) analysis of the 61 soybean HSP70 genes confirmed their stress-inducible expression patterns under both drought and heat stress. These findings provide a thorough overview of the evolution and modification of the GmHSP70 gene family, which will help to determine the functional characteristics of the HSP70 genes in soybean growth and development.

Changes in oleic Acid content of transgenic soybeans by antisense RNA mediated posttranscriptional gene silencing.

The Delta-12 oleate desaturase gene (FAD2-1), which converts oleic acid into linoleic acid, is the key enzyme determining the fatty acid composition of seed oil. In this study, we inhibited the expression of endogenous Delta-12 oleate desaturase GmFad2-1b gene by using antisense RNA in soybean Williams 82. By employing the soybean cotyledonary-node method, a part of the cDNA of soybean GmFad2-1b 801 bp was cloned for the construction of a pCAMBIA3300 vector under the soybean seed promoter BCSP. Leaf painting, LibertyLink strip, PCR, Southern blot, qRT-PCR, and fatty acid analysis were used to detect the insertion and expression of GmFad2-1b in the transgenic soybean lines. The results indicate that the metabolically engineered plants exhibited a significant increase in oleic acid (up to 51.71%) and a reduction in palmitic acid (to <3%) in their seed oil content. No structural differences were observed between the fatty acids of the transgenic and the nontransgenic oil extracts.

[Correlation of expression of PTEN with AKT phosphorylation level in leukemia cells].

The study was aimed to explore the correlation of expression of pten mRNA and PTEN protein with AKT phosphorylation levels in various types of leukemia and to elucidate its role in the pathogenesis of leukemia so as to provide some evidence for using PI3K/AKT pathway inhibitors in future. 128 de novo leukemia patients were enrolled in this study, including 61 AML cases, 27 ALL cases, 24 CML cases, and 16 CLL cases. 21 volunteers were selected as normal control. The RT-PCR and Western blot were used to assay the expressions of pten mRNA, PTEN protein, and P-AKT protein in Jurkat cells, bone marrow mononuclear cells of patients respectively. The results showed that the expressions of pten mRNA and PTEN protein in Jurkat cells were lower than that in normal control group; the expression of pten mRNA in AML group was lower than that in normal control group, but the difference was not significant (p=0.274); the expressions of pten mRNA in ALL, CML, CLL each group were lower than that in normal control group, and the difference was significant (p<0.05). Compared with normal control group, the expression of PTEN protein was lower and the expression of P-AKT protein was higher in AML, ALL, CML, CLL each group, the difference were significant (p<0.05). It is concluded that the a lower expression of PTEN protein and higher expression of P-AKT protein may play an important role on leukemia pathogenesis, and inactivation of PTEN protein mainly occurs in the level of protein translation.