BMSCs attenuate hepatic fibrosis in autoimmune hepatitis through regulation of LMO7-AP1-TGFß signaling pathway.

OBJECTIVE: In a previous study, we reported that transplantation of bone mesenchymal stem cells (BMSCs) significantly attenuated liver damage in a mouse autoimmune hepatitis (AIH) model. Moreover, expression of the LIM domain protein, LMO7, correlated positively with the invasive capacity of hepatoma cells. However, whether LMO7 plays a role in inflammation and fibrosis of AIH remains unknown. This investigation aimed to explore the effect of BMSC transplantation on LMO7 and the role of LMO7 in hepatic fibrosis. MATERIALS AND METHODS: S100-induced murine AIH and LPS-induced hepatocyte injury models were successfully established. Three doses of BMSCs were injected into AIH mice via the tail vein. LPS-treated AML12 cells were co-cultured with BMSCs in vitro. Small interfering (si) LMO7 RNA and T5224 (a specific inhibitor of AP-1) were used to demonstrate the relationship between LMO7-AP1-transforming growth factor (TGF)-ß. RESULTS: Pathological examination and serum alanine and aspartate aminotransferase levels indicated that liver damage was notably ameliorated in the BMSC-treated mice. LMO7 level was upregulated, while AP-1 and TGF-ß levels were downregulated upon intervention with BMSCs. AP-1 expression was upregulated in the siLMO7 group, whereas TGF-ß level was downregulated in the T5224 group when compared to those in the control group. CONCLUSIONS: BMSC transplantation significantly limits liver fibrosis and upregulates the expression of LMO7. LMO7 inhibits the TGF-ß pathway by inhibiting AP-1. This implies that BMSCs are a potential means of treating liver fibrosis. This approach has important implications for the treatment of AIH and other fibrotic diseases.

MNX1-AS1 accelerates the epithelial-mesenchymal transition in osteosarcoma cells by activating MNX1 as a functional oncogene.

OBJECTIVE: To investigate whether MNX1-AS1 can accelerate epithelial-mesenchymal transition (EMT) of osteosarcoma cells via activating MNX1. PATIENTS AND METHODS: The expression pattern of MNX1-AS1 in osteosarcoma tissues and cell lines was examined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Moreover, the cytoplasmic and nuclear levels of MNX1-AS1 in osteosarcoma cells were also determined. The regulatory effects of MNX1-AS1 on viability, clonality, migratory, and invasive abilities of the osteosarcoma cells were evaluated. The relative levels of MNX1 and EMT-related genes influenced by MNX1-AS1 were detected. The methylation ability in the promoter of the osteosarcoma cells transfected with si-MNX1-AS1 or MNX1-AS1 vector was determined by the whole genome bisulfite sequencing. RESULTS: MNX1-AS1 was upregulated in osteosarcoma tissues and cell lines, which was mainly expressed in the nucleus. The knockdown of MNX1-AS1 markedly attenuated viability, clonality, migratory, and the invasive abilities of the osteosarcoma cells. Besides, the transfection of si-MNX1-AS1 in U2OS and MG63 cells downregulated MNX1 and Snail, and upregulated E-cadherin. The methylation ability increased after the knockdown of MNX1-AS1, while the overexpression of MNX1-AS1 obtained the opposite trends. CONCLUSIONS: MNX1-AS1 mediates EMT of the osteosarcoma cells via activating MNX1, thereafter accelerating the progression of the osteosarcoma.

Microarray based analysis of gene expression patterns in pancreatic neuroendocrine tumors.

OBJECTIVE: Pancreatic neuroendocrine tumors (PanNETs) are a small subgroup of tumors with a variety of biological behaviors. MATERIALS AND METHODS: We sought to identify the specially expressed genes and characterize significant pathways in PanNETs compared with non-neoplastic samples. Gene expression profile GSE43795 was obtained from Gene Expression Omnibus database, which included 6 PanNETs and 5 non-neoplastic samples. The differentially expressed genes (DEGs) were identified using Limma package. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were used to enrich the functions and pathways of DEGs. Transcription factors (TFs) and tumor-associated genes (TAGs) were also identified. Finally, a protein-protein interaction (PPI) network was constructed, and hub proteins and functional module were screened out. RESULTS: Total of 821 DEGs (421 down-regulated, 400 up-regulated) were selected. GO and KEGG enrichment analyses showed that up-regulated DEGs were related to several pathways, including type 2 diabetes mellitus, Ca2+ signaling pathway, long-term potentiation, and long-term depression pathways. Down-regulated DEGs were enriched in several pathways, such as pancreatic secretion, protein digestion and absorption, and metabolic pathway. Interferon-stimulated gene protein 15 (ISG15), somatostatin (SST), and synaptosomal-associated protein 25 kDa (SNAP25) were identified as hub proteins. CONCLUSIONS: The genes involved in type 2 diabetes mellitus pathway may play important roles in the development of PanNETs. SNAP25, SST, and ISG15 may be used as potential targets for treatment of PanNETs.

Multiplexed immunoglobulin E sensitization in relation to exhaled nitric oxide in a population sample of children.

This study investigated the relationship between the specific immunoglobulin E (IgE) profile for 40 allergens using a novel microarray technique (BioIC) and fraction of exhaled nitric oxide (FeNO) in a population sample of 1321 children. Significant positive associations were found between FeNO and sensitization to mites (P < 0.001), animals (P = 0.001), cockroaches (P < 0.001), and foods (P = 0.042), and furthermore, between FeNO and the number of sensitizations (all P < 0.05) or the sum of specific IgE (all P ≤ 0.01) against the aforementioned allergen categories. Specifically, sensitization to the following allergens was significantly related to higher FeNO: Dermatophagoides pteronyssinus, Dermatophagoides farina, Blomia tropicalis, cat, German cockroach, Oriental cockroach, codfish, crab, shrimp, and cheese (all P ≤ 0.01). In conclusion, IgE sensitization to mites, pets, cockroaches, seafood, and cheese, respectively, is significantly associated with elevated FeNO levels in a dose-dependent fashion in children. Our results provide new evidence that sensitization to certain food allergens may contribute to prompt inflammation in the airways.

First Report of Fusarium oxysporum Causing Wilt of Dendrobium candidum in Zhejiang Province, China.

White Dendrobium (Dendrobium candidum Wall. ex Lindl.) is a traditional Chinese medicinal herb that is used raw or processed for health care products in China (4). It is propagated by tissue-cultured seedlings largely because of its dramatic decrease in the wild from heavy market demand. In July 2011, a wilt disease was observed on 1-year-old seedlings in a cultivated field in Yueqing City, Zhejiang Province, China. Initial symptoms were one to several chlorotic leaves in a cluster of plants. As the disease progressed, leaves wilted before dropping. Vascular discoloration was not observed in infected stems. Individual plants wilted, bent, and rotted starting from the base and eventually all plants in a cluster wilted. Three strains isolated from stem tissues collected at different sites were grown on potato dextrose agar (PDA) and carnation leaf agar (CLA) under a 12-h alternating cycle of light and dark at a temperature of 25°C. Macroconidia were sparse, three-septate, slightly curved, and ranged from (21.1) 23.9 to 32.7 (36.9) × (2.8) 3.3 to 4.2 (4.8) µm. Microconidia were abundant, unicellular, oval to reniform, and ranged from (5.3) 6.0 to 8.2 (9.4) × (2.2) 2.5 to 3.3 (3.6) µm on PDA. Abundant chlamydospores formed after 3 weeks on CLA. Chlamydospores were single and sometimes in pairs, terminal and intercalary, rough walled, and measured (6.7) 7.6 to 10.2 (11.9) × (5.9) 6.8 to 8.8 (10.4) µm. Strains were identified as Fusarium oxysporum on the basis of morphological features (2). PCR amplification of genomic DNA using the primers ITS6 and ITS4 as well as ef1 and ef2 (translation elongation factor 1-α gene) generated sequences of approximately 500 bp and 670 bp, respectively. ITS sequences were submitted to the NCBI database (Accession Nos. JQ809654, JQ809655, and JQ809656), and were identical to the sequence for F. oxysporum, Accession No. HQ379654 (3). Three sequences of a portion of TEF-1α (Accession Nos. JQ809657, JQ809658, and JQ809659) had 99% similarity with an isolate of F. oxysporum isolate from chickpea (Accession No. HQ731057) (1), thus confirming the identity of the pathogen. The roots of five to seven transplanted seedlings of a cluster grown for 2 months in pasteurized potting media (primarily chipped pine bark) were inoculated with 15 ml of a 107 conidia ml-1 suspension of each isolate. There were 10 replicates for each isolate treatment. Sterile water was used as a negative control. Plants were grown in the greenhouse at 28 ± 2°C. Lateral roots turned brown 15 days after inoculation. Leaves started to turn yellow 1 month after inoculation and the plants began to wilt 2 months after inoculation. Noninoculated controls did not develop disease symptoms. The pathogenic isolates were reisolated from all diseased plants. Isolates have been deposited at the Biotechnology Institute, Zhejiang University (F_01, F_02 and F_03). To the best of our knowledge, this is first record of Fusarium wilt of D. candidum in China. References: (1) D. Jiménez-Fernández et al. Plant Dis. 95:860, 2011. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (3) N. Rajmohan et al. Plant Pathol. 131:631, 2011. (4) J. Z. Zhang et al. Plant Pathol. 57:370, 2008.