Identification and expression analysis of the YABBY gene family in strawberry / 生物工程学报

YABBY proteins are important transcription factors that regulate morphogenesis and organ development in plants. In order to study the YABBY of strawberry, bioinformatic technique were used to identify the YABBY gene families in Fragaria vesca (diploid) and Fragaria×ananassa (octoploid), and then analyze the sequence characters, phylogeny and collinearity of the family members. The RNA-seq data and the quantitative reverse transcription-polymerase chain reaction (qRT-PCR) technique were used to assay the expression patterns of the family members. A green fluorescent protein (GFP) was fused with FvYABBYs and transiently expressed in tobacco leaf cells for the subcellular localization. As the results, six FvYABBY genes and 26 FxaYABBY genes were identified from F. vesca and F.×ananassa, respectively. The FvYABBY genes were grouped into five clades, and five family members were orthologous with AtYABBY genes of Arabidopsis. In F. vesca, all of the FvYABBYs were basically not expressed not expressed in root and receptacle, while FvYABBY1, FvYABBY2, FvYABBY5 and FvYABBY6 were highly expressed in leaf, shoot, flower and achene. In F.×ananassa, FxaYABBY1, FxaYABBY2, FxaYABBY5 and FxaYABBY6 were expressed in achene, and all FxaYABBY were poorly or not expressed in receptacle. Additionally, under the abiotic stresses of low temperature, high salt and drought, the expression of FvYABBY1, FvYABBY3, FvYABBY4 and FvYABBY6 were down-regulated, FvYABBY5 was up-regulated, and FvYABBY2 was up-regulated and then down-regulated. In tobacco leaf cells, the subcellular localization of FvYABBY proteins were in the nucleus. These results provides a foundation for the functional researches of YABBY gene in strawberry.

Genome-wide identification of SUN gene family in Fragaria vesca and stresses-response analysis / 生物工程学报

SUN gene is a group of key genes regulating plant growth and development. Here, SUN gene families of strawberry were identified from the genome of the diploid Fragaria vesca, and their physicochemical properties, genes structure, evolution and genes expression were also analyzed. Our results showed that there were thirty-one FvSUN genes in F. vesca and the FvSUNs encoded proteins were classified into seven groups, and the members in the same group showed high similarity in gene structures and conservative motifs. The electronic subcellular localization of FvSUNs was mainly in the nucleus. Collinearity analysis showed that the members of FvSUN gene family were mainly expanded by segmental duplication in F. vesca, and Arabidopsis and F. vesca shared twenty-three pairs of orthologous SUN genes. According to the expression pattern in different tissues shown by the transcriptome data of F. vesca, the FvSUNs gene can be divided into three types: (1) expressed in nearly all tissues, (2) hardly expressed in any tissues, and (3) expressed in special tissues. The gene expression pattern of FvSUNs was further verified by quantitative real-time polymerase chain reaction (qRT-PCR). Additionally, the seedlings of F. vesca were treated by different abiotic stresses, and the expression level of 31 FvSUNs genes were assayed by qRT-PCR. The expression of most of the tested genes was induced by cold, high salt or drought stress. Our studies may facilitate revealing the biological function and molecular mechanism of SUN genes in strawberry.

miR-182 inhibits proliferation, invasion of endometrial glandular epithelial cells in endometriosis micevia Wnt signaling pathway by targeting AQP5 / 中华内分泌外科杂志

Objective:To elucidate the effect of miR-182 on proliferation, invasionof endometrial glandular epithelial cells in endometrrosis (EMs) mice via Wnt signaling pathway through targeting Aquaporin5 (AQP5) .Methods:The mice model of EMs were established. Subsequently the adenoepithelial tissue of endometrium were collected and the expression of miR-182 and AQP5 in tissue was detected. Enzyme-linked immunosorbent assay (ELISA) was performed to test the expression of inflammatory factors in normal and EMs mice. Then endometrial glandular epithelial cells in mice were isolated and divided into different groups. qRT-PCR and Western blot was performed to detect the expression of miR-182, AQP5,and Wnt pathway related factors (Wnt-1, β-catenin) in cells. The proliferation activity and invasion ability in each group of cells were examined by MTT and Transwell.Results:Compared with Normal mice, the expression of miR-182 was decreased while AQP5 and Wnt pathway related factors (Wnt-1, β-catenin) expression were increased in endometrial glandular epithelial tissues of EMs mice (all P<0.05) . In cell experiments, miR-182 overexpression or AQP5 silencing could inhibit the expression of Wnt pathway related factors (Wnt-1, β-catenin) . At the same time, cell viability as well as invasion ability were decreased (all P<0.05) . Indexes in miR-182 inhibitor group exhibited an opposite trend compared with that in miR-182 mimic group. The effects of sh-AQP5 on EMs cells could be offset by miR-182 inhibitor. Conclusion:Up-regulated expression of miR-182 can reduce the proliferation and invasion of endometrial glandular epithelial cells of EMs mice through inhibiting the activation of Wnt signaling pathway by down-regulating AQP5.

Emodin reduces FFAs-induced fatty degeneration in HepG2 cells via the AMPK/SREBP-1 pathway / 中国医师杂志

Objective To investigate the effects of emodin on the triglyceride metabolism and oxidative stress in steatosis in HepG2 cells and possible underlying mechanisms.Methods The appropriate concentration of emodin on HepG2 cells were detected by methyl thiazolyl tetrazolium (MTT) assay.HepG2 cells were induced to fat overaccumulation by 1 mmol/L free fatty acids (FFA) (oleate∶ palmitate =2∶1).The model group exposed to 10 μmol/L,20 μmol/L,40 μmol/L emodin.The intracellular lipid accumulation was documented by Oil Red O staining and the content of triglyceride and total cholesterol was observed.Reactive oxygen species (ROS) was determined by flow cytometry.Western blotting was performed to analyze the protein levels of adenosine monophosphate-activated protein kinase (AMPK),phosphorylated AMPK,and sterol regulatory element-binding protein 1 (SREBP-1).Results Emodin reduced lipid accumulation and triglycerides (TG) content (P ＜ 0.05).At the same time,it significantly reduced ROS production (P ＜ 0.05).Moreover,the levels of AMPK and p-AMPK protein were significantly upregulated,and SREBP-1 protein was significantly downregulated with the treatment of emodin (P ＜ 0.01).Conclusions This study has demonstrated that emodin can reduce fatty degeneration induced by FFAs in hepatocytes,and this effect may be partially mediated by the AMPK/SREBP-1 pathway.

Effect of viral factors and host cellular immunity on the response to interferon in the patients with chronic hepatitis C / 中华传染病杂志

Objective To determine the influence of viral factors and host cellular immunity on the response to interferon in the patients with chronic hepatitis C. Methods Forty patients with chronic hepatitis C were treated with interferon ?. The relationships between response to interferon a and HCV genotype, quasispecies heterogeneity, HCV RNA level and HCV specific cytotoxic T lymphocyte (HCV CTL) activity in the liver were analyzed. Results After 6 months of therapy, 21 patients had obtained end of treatment response (ETR), 10 Patients of which had obtained sustained response (SR). The other 19 patients got no response (NR). ETR rate in patients with genotype HCV1 infection (43.3%, 13/30) was significantly lower than that in patients with non HIV1 infection (80%, 8/10) [ P

Activity of HCV-specific cytotoxic T lymphocytes in chronic hepatitis C / 中华传染病杂志

Objective To define the role of HCV-specific cytotoxic T lymphocytes (CTL) in chro-nic hepatitis C virus (HCV) infection. Methods HCV-specific CTL activity (HCV-CTL) was assessed in the liver and peripheral blood cells in 62 patients with chronic hepatitis C and 8 non-HCV-infection controls by using bulk-expanded liver/peripheral blood mononuclear lymphocytes (PBMC)-derived CD8+ cells as effector cells, EBV-transformed B cells as autologous target cells and recombinant vaccinia vectors expressing various regions of the HCV genome as transduction vector, in a standard chromium release assay. Results HCV-CTL activity was detected from the liver in 28 of the 60 patients (46.7%), but not from PBMC. CTL activity could not be detected from the liver and PBMC in all non-HCV-infection controls. Five patients with non-type 1 HCV infection were found to have HCV-specific CTL activity against HCV type 1 epitope. Compared with the patients without detectable HCV-specific CTL activity based on our assay, those exhibiting CTL activity had higher serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, higher histologic activity indices and lower levels of HCV RNA (all P