Effect of Bcl-2-siRNA on proliferation and apoptosis of pediatric acute B lymphoblastic leukemia (A-BLL) cells.

This study analyzed the effect of small interfering RNA specific for the Bcl-2 gene (siRNA Bcl-2) on the proliferation and chemotherapeutic sensitivity of pediatric A-BLL cells. Marrow samples were obtained from sixty newly-diagnosed A-BLL pediatric patients. The Bcl-2 mRNA expression in these samples was quantified by real time polymerase chain reaction. The Bcl-2 mRNA re-expression was analyzed by RNA interference using Bcl-2-siRNA. Cellular proliferation was detected using the MTT (Thiazolyl Blue Tetrazolium Bromide) assay. The cell apoptosis was quantified by flow cytometry. The Bcl-2 mRNA expression was significantly higher in the drug-resistance group than in the chemotherapy sensitivity group prior to chemotherapy (P < 0.05). In addition, the Bcl-2 mRNA expression in the chemotherapy sensitivity group was significantly higher before chemotherapy than that after chemotherapy (P < 0.05). The Bcl-2 mRNA expression significantly decreased in the leukemic cells of the Bcl-2-siRNA transfection group. We observed statistically significant differences in the relative mRNA expression levels among the Bcl-2-siRNA transfection, blank control, liposome empty transfection, and unrelated sequence oligonucleotide groups (P < 0.05). The rate of apoptosis in pediatric A-BLL leukemic cells was observed to increase significantly after transfection with Bcl-2-siRNA compared to the control, liposome empty transfection, and unrelated sequence oligonucleotide groups (P < 0.05). Therefore, we concluded that Bcl-2-siRNA can successfully inhibit the multiplicative capacity of A-BLL leukemic cells and promote apoptosis.

Induction and identification of Stylosanthes guianensis tetraploids.

Stylosanthes guianensis is an elite and important forage legume species, which is extensively cultivated in tropical areas. Polyploid breeding via exposure to colchicine is a conventional and practical method to improve varieties of S. guianensis. Terminal buds of S. guianensis Reyan No.5 seedlings were treated with different concentrations of colchicine (0.00, 0.05, 0.10, 0.15, 0.20, and 0.25%) for 24, 48, and 72 h. Morphological and cytological variants were observed at a frequency of <96% among transplanted seedlings. The cytogenetic analysis of young leaf cells was conducted on all variants to identify their ploidy levels. The most efficient procedure for tetraploid production was the treatment of seedling apical buds with 20% colchicine for 48 h, with the tetraploid induction rate being 10%. This is a relatively simple and reliable method for the production of tetraploidy in S. guianensis.

Physical localization of molecular markers and assignment of the 15th linkage group to chromosome 11 of the karyotype in cassava (Manihot esculenta Crantz) by primed in situ labeling.

Physical localization of molecular markers and assignment of the 15th linkage group to chromosome 11 of the karyotype in cassava (Manihot esculenta Crantz) were achieved using primed in situ labeling. Amplified signals for both the EST507-1 and SSRY13-5 markers were consistently observed in different stages of cell division. A comparison of the length, arm ratio, and other morphological characteristics of somatic metaphase chromosomes in karyotype analysis indicated that the EST507-1 and SSRY13-5 markers were localized on the short and long arm of cassava chromosome 11 with the relative map positions of 41.67 and 23.07, respectively. The physical localization of the 2 markers on chromosome 11 of the karyotype corresponds to their positions on the 15th linkage group in cassava.

Disease indicators for sepsis and analysis of sepsis treatment in children using the continuous blood purification technique.

We analyzed disease severity, inflammation markers, and dynamic changes in cartilage glycoprotein 39 (YKL-40) and C-reactive protein (CRP) levels in children with sepsis before and after treatment with continuous blood purification (CBP). Study participants were 30 children with severe sepsis who were cured from the disease (experimental group) in the Children's Serious Disease Center of In-ner Mongolia People's Hospital between June 2012 and October 2013. Symptomatic CBP treatment was performed after disease severity scoring. Interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), YKL-40, and CRP levels were tested 0, 12, 24, and 48 h after CBP treatment. YKL-40 mRNA expression in whole blood was determined biochemically, and its expression in peripheral blood was determined with an immunochemical method. We found a significant difference in disease severity scores before and 48 h after CBP treatment (P < 0.05). IL-6, TNF-α, YKL-40, and CRP levels in children with sepsis at 12, 24, and 48 h after CBP treatment significantly differed from those before treatment (P < 0.05). The relative expression of YKL-40 mRNA in the experimental group before CBP treatment significantly increased from that of the control group (P < 0.05). We found a positive correlation between IL-6, TNF-α, YKL-40, and CRP levels 48 h after CBP treatment. In conclusion, CBP is an effective treatment strategy for pediatric sepsis. YKL-40 and CRP can be used to evaluate the effects of sepsis treatment.

Influence of thoracic duct ligation on the lipid metabolism of patients with esophageal carcinoma after esophagectomy.

The aim of this study was to determine the influence of thoracic duct ligation on the lipid metabolism of patients with esophageal carcinoma after esophagectomy. A total of 74 patients with esophageal carcinoma who underwent esophagectomy were divided into 2 groups according to whether or not their thoracic duct was ligated. Thirty-nine patients were in the thoracic duct ligation group and the other 35 assigned to the control group. Enteral feeding was through a nasojejunal tube from the 1st day to the 8th day after operation, and liquid diet was provided starting on the 6th day. We compared the plasma concentrations of cholesterol (CHOL), triglycerides (TG), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) at different time points. There were no statistically significant differences between the two groups in CHOL, TG and HDL levels at different times. However, LDL levels in the thoracic duct ligation group were significantly lower at different times compared to the other group (P < 0.05), where they were the lowest at the end of the 1st month and then gradually recovered 3 months later. Thoracic duct ligation can effectively prevent chylomicrons from being transferred to the blood, reducing the generation of LDL. The establishment of collateral circulation was slow after the ligation of the thoracic duct, which had a negative effect on early postoperative nutrition of patients.

Isolation of 1-aminocyclopropane-1-carboxylate synthase gene from Oncidium Gower Ramsey.

A full-length cDNA of a 1-aminocyclopropane-1-carboxylate synthase (ACS) family member from Oncidium, named OnACS1 (GenBank accession No. JQ822087) was cloned and characterized by reverse transcription polymerase chain reaction and rapid amplification of cDNA ends technology. The full-length cDNA was 1586 bp, including a 1308-bp open reading frame, a 105-bp 5' untranslated region (UTR), and 173-bp 3' UTR, encoding 436 amino acids. The deduced amino acid sequence of OnACS1 shares 85, 84, and 83% homology with ACS proteins of Cattleya bicolor, Dendrobium crumenatum, and Phalaenopsis hybrid, respectively. Prokaryotic expression and sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that a specific band was produced and was consistent with the predicted protein size. A tissue-specific manner of OnACS1 expression was observed, and it was predominantly expressed in the gynostemium. The OnACS1 expression in the sepals and gynandria was upregulated by 1% ethephon treatment.

Effect of MPO/H2O2/NO(-) system on nitric oxide-mediated modification of TTR amyloid and serum TTR in FAP ATTR Val30Met patients.

Amyloid deposits consist of protein fibrils and amorphous material, and this deposition is related to oxidative stress. Previously, we demonstrated the presence of high-density lipoproteins and/or lipids in amyloid deposits of familial amyloid polyneuropathy patients. In this study, the presence of myeloperoxidase (MPO) in amyloid deposits was demonstrated using immunohistochemical staining. In contrast, normal surrounding tissues were consistently negative for MPO. Nitrotyrosine was present in amyloid deposits after being exposed to the MPO/H2O2/NO(-) system by immunohistochemical staining, and the oxide mediated modification of serum transthyretin (TTR) was observed upon exposure to the MPO/H2O2 system using two-dimensional gel electrophoresis and TTR Western blotting. This observation revealed that the TTR amyloid deposits and serum TTR were oxidized by the MPO/H2O2/NO(-) system. Nitric oxide-mediated modification of TTR may play a role in amyloidogenesis in vivo.

Evaluation of genetic diversity in Piper spp using RAPD and SRAP markers.

Random amplified polymorphic DNA (RAPD) and sequence-related amplified polymorphism (SRAP) analysis were applied to 74 individual plants of Piper spp in Hainan Island. The results showed that the SRAP technique may be more informative and more efficient and effective for studying genetic diversity of Piper spp than the RAPD technique. The overall level of genetic diversity among Piper spp in Hainan was relatively high, with the mean Shannon diversity index being 0.2822 and 0.2909, and the mean Nei's genetic diversity being 0.1880 and 0.1947, calculated with RAPD and SRAP data, respectively. The ranges of the genetic similarity coefficient were 0.486-0.991 and 0.520-1.000 for 74 individual plants of Piper spp (the mean genetic distance was 0.505 and 0.480) and the within-species genetic distance ranged from 0.063 to 0.291 and from 0.096 to 0.234, estimated with RAPD and SRAP data, respectively. These genetic indices indicated that these species are closely related genetically. The dendrogram generated with the RAPD markers was topologically different from the dendrogram based on SRAP markers, but the SRAP technique clearly distinguished all Piper spp from each other. Evaluation of genetic variation levels of six populations showed that the effective number of alleles, Nei's gene diversity and the Shannon information index within Jianfengling and Diaoluoshan populations are higher than those elsewhere; consequently conservation of wild resources of Piper in these two regions should have priority.