Knockdown the moyamoya disease susceptibility gene, RNF213, upregulates the expression of basic fibroblast growth factor and matrix metalloproteinase-9 in bone marrow derived mesenchymal stem cells.

Moyamoya disease (MMD) is a chronic, progressive cerebrovascular occlusive disease. Ring finger protein 213 (RNF213) is a susceptibility gene of MMD. Previous studies have shown that the expression levels of angiogenic factors increase in MMD patients, but the relationship between the susceptibility gene RNF213 and these angiogenic mediators is still unclear. The aim of the present study was to investigate the pathogenesis of MMD by examining the effect of RNF213 gene knockdown on the expression of matrix metalloproteinase-9 (MMP-9) and basic fibroblast growth factor (bFGF) in rat bone marrow-derived mesenchymal stem cells (rBMSCs). Firstly, 40 patients with MMD and 40 age-matched normal individuals (as the control group) were enrolled in the present study to detect the levels of MMP-9 and bFGF in serum by ELISA. Secondly, Sprague-Dawley male rat BMSCs were isolated and cultured using the whole bone marrow adhesion method, and subsequent phenotypic analysis was performed by flow cytometry. Alizarin red and oil red O staining methods were used to identify osteogenic and adipogenic differentiation, respectively. Finally, third generation rBMSCs were transfected with lentivirus recombinant plasmid to knockout expression of the RNF213 gene. After successful transfection was confirmed by reverse transcription-quantitative PCR and fluorescence imaging, the expression levels of bFGF and MMP-9 mRNA in rBMSCs and the levels of bFGF and MMP-9 protein in the supernatant of the culture medium were detected on the 7th and 14th days after transfection. There was no significant difference in the relative expression level of bFGF among the three groups on the 7th day. For the relative expression level of MMP-9, there were significant differences on the 7th day and 14th day. In addition, there was no statistically significant difference in the expression of bFGF in the supernatant of the RNF213 shRNA group culture medium, while there was a significant difference in the expression level of MMP-9. The knockdown of the RNF213 gene affects the expression of bFGF and MMP-9. However, further studies are needed to determine how they participate in the pathogenesis of MMD. The findings of the present study provide a theoretical basis for clarifying the pathogenesis and clinical treatment of MMD.

Role of dietary tea polyphenols on growth performance and gut health benefits in juvenile hybrid sturgeon (Acipenser baerii ♀ × A. schrenckii ♂).

The present study aimed to evaluate the effects of dietary TPs on growth performance, intestinal digestion, microflora and immunity in juvenile hybrid sturgeon. A total of 450 fish (97.20 ± 0.18 g) were randomly divided into a standard diet (TP-0) or four treatments consisting of a standard diet supplemented with four concentrations of TPs (mg/kg): 100 (TP-100), 300 (TP-300), 500 (TP-500), and 1000 (TP-1000) for 56 days. The TP-300 significantly increased weight gain rate (WGR) and specific growth rate (SGR) (p < 0.05), and TP-1000 significantly increased the feed conversion ratio (FCR) (p < 0.05). TP-300 and TP-500 significantly increased intestinal trypsin, amylase, and lipase activities (p < 0.05). Besides, TP-300 significantly enhanced total antioxidant capacity (T-AOC) and the levels of superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) and decreased malondialdehyde (MDA) content (p < 0.05). Moreover, TP-300 decreased the expression levels of tumor necrosis factor-alpha (TNF-α), interleukin 8 (IL-8), and interleukin 1ß(IL-1ß) compared with TP-0 and TP-1000 (p < 0.05). In addition, the intestinal microbiota diversity in the TP-300 group was observably higher, the dominant microbiota was Bacteroidota, Cyanobacteria, Proteobacteria and Firmicutes at the phylum level, Enterobacteriaceae, Nostocaceae and Clostridiaceae at the family level. The relative abundances of potential probiotics including Rhodobacteraceae and potential pathogens especially Clostridiaceae were the highest, and lowest, respectively. In conclusion, TP-300 altered the abundance of microbial taxa, resulting in enhancing the intestinal digestion, antioxidant status and non-specific immunity to improve the growth performance in juvenile hybrid sturgeon.

Stress responses of the intestinal digestion, antioxidant status, microbiota and non-specific immunity in Songpu mirror carp (Cyprinus carpio L.) under starvation.

Songpu mirror carp, Cyprinus carpio L., is a new variety of common carp that has become an economically important freshwater fish in China. However, it remains unknown how its metabolism is regulated under starvation. Here, we investigated how intestinal digestion, antioxidant status, microbiota and immune activities were affected under starvation stress. The feeding regimes were designed as follows: ST0 comprised fish allowed to feed continuously; ST1 comprised fish starved for 1 week; ST2 comprised fish starved for 2 weeks; ST3 comprised fish starved for 3 weeks; ST4 comprised fish starved for 4 weeks. Our results showed a significant decrease in the level of intestinal amylase, lipase, and protease activities in the group ST4 (P < 0.05). Compared with the control group, intestinal antioxidant enzyme activities were significantly increased during short-term starvation. The gene expression levels of interleukin 1ß (IL-1ß), interleukin 8 (IL-8) and tumor necrosis factor-alpha (TNF-α) were elevated in the groups ST3 and ST4. We also detected the reduction in the expression levels of interleukin 10 (IL-10) and transforming growth factor ß (TGF-ß2) compared with those of the group ST0. Notably, the gut microbial composition was dominated by Proteobacteria, Acidobacteria, Actinobacteria, Bacteroidetes, and Firmicutes. The relative abundance of the dominant microbial phyla changed significantly under starvation stress. Taken together, our results suggest that starvation can induce the change of intestinal digestion, non-specific immunity and microbiota in Songpu mirror carp, and provide new insights into its habitat selection and adaptation to environmental changes.

[Treatment of moyamoya disease by superficial temporal artery-middle cerebral artery anastomosis].

OBJECTIVE: To explore the clinical efficacy and experiences of superficial temporal artery-middle cerebral artery anastomosis (STA-MCA) for the treatment of adult Moyamoya disease. METHOD: Preoperative and postoperative clinical data of 60 patients with moyamoya disease from affiliated hospital of jining medical college were analyzed retrospectively, including 46 cases of ischemic lesion and 14 cases of hemorrhage lesion. All patients were diagnosed with moyamoya disease by DSA and received STA-MCA anastomosis. RESULTS: The clinical symptoms of 52 patients disappeared after surgery; the clinical symptoms were improved in 6 patients; 1 patient complicated with cerebral infarction; 1 patients reoccurred cerebral hemorrhage broken into ventricles 1 year after anastomosis. CONCLUSION: STA-MCA anastomosis can improve the symptoms of ischemic moyamoya disease effectively and reduce the recurrence of cerebral hemorrhage. STA-MCA anastomosis is a rational and effective approach for the treatment of adult Moyamoya disease.

RNF213 gene silencing upregulates transforming growth factor ß1 expression in bone marrow-derived mesenchymal stem cells and is involved in the onset of Moyamoya disease.

Moyamoya disease (MMD) is a chronic and progressive cerebrovascular occlusion disease, the precise etiology of which is poorly understood. Ring finger protein 213 (RNF213) has been previously identified as a susceptibility gene that serves an important role in angiogenesis, where it has been shown to be closely associated with the onset of MMD. Patients with MMD exhibit increased expression levels of various pro-inflammatory molecules and angiogenic factors. Under certain conditions, bone marrow mesenchymal stem cells (BMSCs) have the ability to differentiate to form neuron-like and microglia-like cells. In the present study, a total of 40 MMD patients and 40 healthy individuals were enrolled. ELISA assays revealed that the expression of serum vascular endothelial growth factor (VEGF) and transforming growth factor ß1 (TGF-ß1) were higher than that in healthy controls. Furthermore, rat BMSCs (rBMSCs) were isolated and cultured using the whole bone marrow adherence method, which were then phenotyped using flow cytometry. Osteogenic and adipogenic differentiation were determined by using Alizarin red and oil red O staining, respectively. RNF213 was knocked-down using a lentivirus-mediated short hairpin RNA system in passage three rBMSCs, and successful transfection of the RNF213 was confirmed by RT-qPCR and fluorescence imaging. The expression levels of VEGF and TGF-ß1 in these rBMSCs were measured on days 7 and 14, respectively. The results demonstrated that RNF213 knockdown upregulated TGF-ß1 at both protein and mRNA levels, but did not exert any effect on VEGF gene expression. In conclusion, these findings suggested that that RNF213 knockdown may contribute to aberrant TGF-ß1 expression via a pathway that remains to be unidentified, indicating that quantitative changes in RNF213 gene expression may serve an important role in the pathogenesis of MMD.

Complete genome sequence data of multidrug-resistant Stenotrophomonas sp. strain SXG-1.

Objectives A multidrug-resistant bacterium, Stenotrophomonas sp. SXG-1, was isolated from the liver of diseased hybrid sturgeon from Guizhou province, China. Methods Whole-genome sequencing was performed on the Illumina HiSeq 2500-PE125 platform with MPS (massively parallel sequencing) Illumina technology. All good quality paired reads were assembled using the SOAPdenovo into a number of scaffolds. PHI (Pathogen Host Interactions), VFDB (Virulence Factors of Pathogenic Bacteria) and ARDB (Antibiotic Resistance Genes Database) were used to analyses pathogenicity and drug resistance. Results Here we reported the complete genome sequence of Stenotrophomonas sp. SXG-1, which comprised 4534,602bp in 4077 coding sequences (CDS) with a G+C content of 66.42%. The genome contained 4 gene islands, 72 tRNAs and 13 rRNAs. According to the annotation analysis, strain SXG-1 encoded 22 genes related to the multidrug resistance. In addition to 10 genes conferring resistance to antimicrobial drugs of different classes via alternative mechanisms, 12 genes of efflux pumps were presented, 9 of which were reported for the first time in Stenotrophomonas maltophilia. Conclusion This was the first complete genome sequence of Stenotrophomonas sp. isolated from the sturgeon. The complete genome sequence of Stenotrophomonas sp. strain SXG-1 may provide insights into the mechanism of antimicrobial resistance and prevent disease.

CpG island methylator phenotype association with elevated serum alpha-fetoprotein level in hepatocellular carcinoma.

PURPOSE: CpG island methylator phenotype (CIMP) involves hypermethylation targeted toward the promoters of multiple genes. To gain insight into the role of epigenetic aberration of tumor-related genes in hepatocarcinogenesis, we determined a hypermethylation profile in hepatocellular carcinoma (HCC). EXPERIMENTAL DESIGN: We examined the promoter methylation status of nine genes in 50 HCCs, 50 paired nontumor tissues, and 6 normal liver tissues by methylation-specific PCR. CIMP+ was defined as having five genes that are concordantly methylated. RESULTS: The frequency of promoter methylation of nine genes in 50 HCCs varied from 10% in P53 to 94% in c-Myc. The methylation status of P14, P15, P16, ER, RASSF1A, WT1, and c-Myc was significantly correlated with HCC and nontumor tissues (P<0.05). Hypermethylation of one or more genes was found in 96% of HCC. CIMP was more frequent in HCC than in nontumor tissues (70% and 12%, P<0.001). There is a significant association between CIMP and methylation of P14, P15, P16, ER, RSAAF1A, and WT1 (P<0.05) and serum alpha-fetoprotein (AFP) level (P=0.017). CIMP+ was more frequent in HCC with AFP>or=30 microg/L than those with AFP<30 microg/L (P=0.005). In addition, the promoter hypermethylation of P15 and P16 was associated with elevated serum AFP levels in 35 HCC samples with CIMP+ (P<0.05). CONCLUSIONS: Positive correlation of CIMP and AFP levels in HCC suggests that CIMP can serve as a molecular marker of late-stage HCC development.

The complete mitochondrial genome sequence of Schizothorax griseus (Cypriniformes: Cyprinidae).

In this study, the complete mitochondrial genome (mtDNA) of Schizothorax griseus was sequenced. The mitogenome is 16,586 bp in length, composed of 13 protein-coding genes, two rRNA genes, 22 tRNA genes, and two non-coding regions. The phylogenetic relationship constructed by Bayesian inference (BI) method showed that schizothorax is not a monophyly, species in the same river express a more close relationship. The results can contribute to explore the high level of diversification and the taxonomic conflict caused by morphologic characters of schizothorax, also, it will be an evidence for the biogeography and evolutionary mechanisms of Schizothoracinae fishes.

Effect of Corilagin on the Proliferation and NF-κB in U251 Glioblastoma Cells and U251 Glioblastoma Stem-Like Cells.

Background. This study is to explore the effect of corilagin on the proliferation and NF-κB signaling pathway in U251 glioblastoma cells and U251 glioblastoma stem-like cells. Methods. CD133 positive U251 glioblastoma cells were separated by immunomagnetic beads to isolate glioblastoma stem-like cells. U251 cells and stem-like cells were intervened by different corilagin concentrations (0, 25, 50, and 100 µg/mL) for 48 h, respectively. Cell morphology, cell counting kit-8 assay, flow cytometry, dual luciferase reporter assay, and a western blot were used to detect and analyze the cell proliferation and cell cycle and investigate the expression of IKBα protein in cytoplasm and NF-κB/p65 in nucleus. Results. Corilagin inhibited the cell proliferation of U251 cells and their stem-like cells and the inhibition role was stronger in U251 stem-like cells (P < 0.05). The cell cycle was arrested at G2/M phase in the U251 cells following corilagin intervention; the proportion of cells in G2/M phase increased as the concentration of corilagin increased (P < 0.05). The U251 stem-like cells were arrested at the S phase following treatment with corilagin; the proportion of cells in the S phase increased as the concentration of corilagin increased (P < 0.05). The ratio of dual luciferase activities of U251 stem-like cells was lower than that of U251 cells in the same corilagin concentration. With increasing concentrations of corilagin, the IKBα expression in cytoplasm of U251 cells and U251 stem-like cells was increased, but the p65 expression in nucleus of U251 cells and U251 stem-like cells was decreased (P < 0.05). Conclusion. Corilagin can inhibit the proliferation of glioblastoma cells and glioblastoma stem-like cells; the inhibition on glioblastoma stem-like cell proliferation is stronger than glioblastoma cells. This different result indicates that the effect of corilagin on U251 cells and U251 stem-like cells may have close relationships with mechanism of cell cycle and NF-κB signaling pathway; however, the real antitumor mechanism of corilagin is not yet clear and requires further study.

p300 expression repression by hypermethylation associated with tumour invasion and metastasis in oesophageal squamous cell carcinoma.

BACKGROUND: Aberrant promoter methylation is an important mechanism for gene silencing. AIMS: To evaluate the promoter methylation status of p300 gene in patients with oesophageal squamous cell carcinoma (OSCC). METHODS: The methylation status of p300 promoter was analysed by methylation-specific PCR (MSP) in 50 OSCC tissues and the matching non-cancerous tissues. Oesophageal cancer cell lines (ECa-109 and TE-10) were treated with the demethylation agent 5-aza-2'-deoxycytidine (5-Aza-CdR), and p300 mRNA expression was detected by RT-PCR. RESULTS: p300 methylation was found in 42% (21/50) of the OSCC tissues, but in only 20% (10/50) of the corresponding non-cancerous tissues (p = 0.017). In OSCC samples, 65% of those with deep tumour invasion (adventitia) and 63% samples with metastasis revealed p300 promoter methylation (p<0.05). p300 mRNA expression was observed in 19.0% (4/21) of methylated tumours and 58.6% (17/29) of unmethylated tumours (p = 0.005). In addition, p300 mRNA expression was observed in 40% (4/10) of methylated non-neoplastic tissues and 87.5% (35/40) of unmethylated non-tumours (p = 0.001). The demethylation caused by 5-Aza-CdR increased the p300 mRNA expression levels in oesophageal cancer cell lines. CONCLUSIONS: p300 transcription silenced by promoter hypermethylation could play a role in the pathogenesis of oesophageal squamous cell carcinoma.