[Identification of GeERF transcription factors in Gelsmium elegans and their expression under low temperature stress].

With prominent medicinal value, Gelsemium elegans has been overexploited, resulting in the reduction of the wild resource. As a result, artificial cultivation turns out to be a solution. However, this medicinal species is intolerant to low temperature, and thus genes responding to the low temperature are important for the cultivation of this species. Based on the transcriptome database of G. elegans at 4 ℃, 29 differentially expressed GeERF genes were identified. Bioinformatics analysis of 21 GeERF gene sequences with intact open reading frames showed that 12 and 9 of the GeERF proteins respectively clustered in DREB subgroup and ERF subgroup. GeDREB1 A-1-GeERF6 B-1, with molecular weight of 23.78-50.96 kDa and length of 212-459 aa, were all predicted to be hydrophilic and in nucleus. Furthermore, the full-length cDNA sequence of GeERF2B-1 was cloned from the leaves of G. elegans. Subcellular localization suggested that GeERF2B-1 was located in the nucleus. According to the quantitative reverse-transcription PCR(qRT-PCR), GeERF2B-1 showed constitutive expression in roots, stems, and leaves of G. elegans, and the expression was the highest in roots. In terms of the response to 4 ℃ treatment, the expression of GeERF2B-1 was significantly higher than that in the control and peaked at 12 h, suggesting a positive response to low temperature. This study lays a scientific basis for the functional study of GeERF transcription factors and provides gene resources for the improvement of stress resistance of G. elegans.

[Effect of aluminum phosphate gel and Kangfuxin on esophageal pathology and IL-8 and PGE2 expressions in a rat model of reflux esophagitis].

OBJECTIVE: To explore the effect of aluminum phosphate gel and Kangfuxin on esophageal pathology and expressions of interleukin-8 (IL-8) and prostaglandin E2 (PGE2) in rats with reflux esophagitis and explore the possible mechanisms. METHODS: Sixty SD rats were randomized into aluminum phosphate gel group (n=10), Kangfuxin group (n=10), aluminum phosphate gel+Kangfuxin group (n=10), model group (n=20), and control group (n=10). Except for those in the control group, all the rats were subjected to infusion of diluted lysolecithin with hydrochloric acid in the esophagus for 14 days. Ten rats in the model group and those in the control group were sacrificed to examine the pathological changes and contents of IL-8 and PGE2 in the esophagus using optical and electron microscopes and radioimmunoassay. The next day the rest rats were given corresponding treatments (saline in model group) administered into the esophagus on a daily basis for 14 days, after which esophageal pathologies and IL-8 and PGE2 contents were examined. RESULTS: The model rats showed obvious esophageal pathologies including inflammatory cell infiltration, vacuolar degeneration of the epithelial cells, esophageal erosion and even ulceration, with severe detachment of the epithelial cells. The rats in all the intervention groups showed lessened esophageal pathologies and lowered esophageal IL-8 and PGE2 contents compared with those in the model group. Esophageal mucosal injury index and IL-8 and PGE2 contents were all significantly lower in rats receiving combined treatment with aluminum phosphate and Kangfuxin than in those receiving either of the treatments (P<0.05). CONCLUSIONS: Both Kangfuxin and aluminum phosphate gel are effective in the treatment for reflux esophagitis induced by lysolecithin and hydrochloric acid, and their therapeutic effects are achieved possibly by reducing IL-8 and PGE2 levels in the esophagus.

Possible role for the thioredoxin system in the protective effects of probucol in the pancreatic islets of diabetic rats.

1. Probucol, a lipid-lowering agent with a potent anti-oxidant action, protects diabetic pancreatic islets by an as yet unknown mechanism. Thioredoxin-interacting protein (TXNIP), an endogenous inhibitor of the ubiquitous thiol oxidoreductase thioredoxin (TRX), has been associated with oxidative stress in diabetic rat islets. The aim of the present study was to examine the effects of probucol on diabetic islet function and expression of TRX and TXNIP. 2. Thirty rats were randomly assigned to one of three groups: a normal control group, a diabetic group and a probucol-treated diabetic group. After 8 weeks treatment with probucol (500 mg/kg per day), plasma malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) activity were determined using chemical colourimetric methods. In addition, the expression of insulin, TRX and TXNIP in islets was analysed using immunohistochemical, western blot and reverse transcription-polymerase chain reaction methods. 3. The expression of TRX and insulin in islets and plasma SOD and CAT activity were lower, but the expression of TXNIP in islets and plasma MDA were higher, in diabetic compared with normal control rats. Upregulated expression of TRX and insulin and downregulated expression of TXNIP were observed in probucol-treated diabetic rats. Probucol treatment increased plasma SOD, decreased plasma MDA and improved hypoinsulinaemia in diabetic rats. 4. The results indicate that treatment with probucol decreases TXNIP expression and increases TRX expression, which may alleviate hypoinsulinaemia by reducing oxidative stress. Therefore, probucol shows promise as a supplemental therapy for islet protection in Type 2 diabetes mellitus.

[Relationship of levels of CD4(+)CD25(+) regulatory T cells and expression of Foxp3 mRNA in peripheral blood with serum immunoglobulin E level in children with bronchiolitis].

OBJECTIVE: To study the roles of CD4(+)CD25(+) regulatory T cells and Foxp3 mRNA in peripheral blood as well as serum total immunoglobulin E (IgE) in the pathogenesis of bronchiolitis caused by respiratory syncytial virus (RSV). METHODS: The proportion of CD4(+)CD25(+) regulatory T cells and expression of Foxp3 mRNA in peripheral blood, and total serum IgE level were tested by flow cytometry, RT-PCR and ELISA respectively in 57 children with RSV bronchiolitis (26 atopic patients and 31 nonatopic patients). Twenty five healthy children were used as the control group. RESULTS: The proportion of CD4(+)CD25(+) regulatory T cells in peripheral blood in children with bronchiolitis, either in the atopic (7.7+/- 1.6%)or the nonatopic group (8.8+/- 2.1%), was significantly lower than that in the control group (10.5+/- 1.6%) (P< 0.01). Foxp3 mRNA expression in peripheral blood was significantly lower in both atopic and nonatopic children with bronchiolitis than that in the control group (P< 0.01). Significantly increased total serum IgE level was noted in both atopic (241.2+/- 102.5 IU/mL) and nonatopic children (125.5+/- 63.2 IU/mL) with bronchiolitis compared with that in the control group (27.2+/- 10.5 IU/ml) (P< 0.01). There were significant differences in the proportion of CD4(+)CD25(+) regulatory T cells and Foxp3 mRNA expression in peripheral blood (P< 0.05) as well as total serum IgE level (P< 0.01) between the atopic and the nonatopic group. The proportion of CD4(+)CD25(+) regulatory T cells (r=-0.70, P< 0.01) and Foxp3 mRNA expression in peripheral blood (r=-0.79, P< 0.01) were closely negatively correlated to total serum IgE level. CONCLUSIONS: Both the proportion of CD4(+)CD25(+) regulatory T cells and Foxp3 mRNA expression in peripheral blood were reduced, in contrast, the total serum IgE level increased in children with RSV bronchiolitis. This suggested that CD4(+)CD25(+) regulatory T cells and Foxp3 mRNA together with IgE participated in the pathogenesis of RSV bronchiolitis.