Basing RNA-seq explored the regulatory mechanism of the carbohydrate metabolism pathways during chicken male germ cell differentiation
IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL
Authors: Lian, Chao; Zuo, Qisheng; Li, Dong; Zhang, Lei; Ahmed, Mahmoud F.; Xiao, Tianrong; Tang, Beibei; Wang, Yingjie; Jin, Kai; Zhang, Yani; Li, Bichun
Abstract
Our study aimed to explore the regulatory mechanism of the carbohydrate metabolism signaling pathways and related genes during the differentiation of chicken embryonic stem cells to male germ cells, providing the basis for improving the efficiency of the in vitro induction system. Cell sorting was used to obtain highly purified embryonic stem cells (ESCs), primitive germ cells (PGCs), and spermatogonial stem cells (SSCs). The total RNA was then extracted from each cell type. The transcriptions of ESCs, PGCs, and SSCs were sequenced by DNA microarray and mRNA sequencing (RNA-seq). The results were analyzed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database. The key pathways and genes of carbohydrate metabolism were screened during the differentiation process of chicken male germ cell. We concluded that 419 differentially expressed genes enriched to 26 carbohydrate metabolism pathways during the differentiation process of ESCs to SSCs, all of the chondroitin sulfate (CS) signaling pathway was significant. We screened the key genes CHSY3, B3GAT1, CHPF, and B4GALT7 which was significantly expressed in CS pathway. Quantitative RT-PCR showed that the expression trend of these genes is consistent with DNA Microarray and RNA-seq results. Our study supports the opinion that CS pathway is significantly different during the differentiation of chicken male germ cell (P < 0.05) and that CHSY3, B3GAT1, CHPF, and B4GALT7 are key genes.
The tumour microenvironment influences survival and time to transformation in follicular lymphoma in the rituximab era
BRITISH JOURNAL OF HAEMATOLOGY
Authors: Blaker, Yngvild Nuvin; Spetalen, Signe; Brodtkorb, Marianne; Lingjaerde, Ole Christian; Beiske, Klaus; Ostenstad, Bjorn; Sander, Birgitta; Wahlin, Bjorn Engelbrekt; Melen, Christopher Michael; Myklebust, June Helen; Holte, Harald; Delabie, Jan; Smeland, Erlend Bremertun
Abstract
The tumour microenvironment influences outcome in patients with follicular lymphoma (FL), but its impact on transformation is less studied. We investigated the prognostic significance of the tumour microenvironment on transformation and survival in FL patients treated in the rituximab era. We examined diagnostic and transformed biopsies from 52 FL patients using antibodies against CD3, CD4, CD8, CD21 (CR2), CD57 (B3GAT1), CD68, FOXP3, TIA1, PD-1 (PDCD1), PD-L1 (CD274) and PAX5. Results were compared with a second cohort of 40 FL patients without signs of transformation during a minimum of five years observation time. Cell numbers and localization were semi-quantitatively assessed. Better developed CD21+ follicular dendritic cell (FDC) meshworks at diagnosis was a negative prognostic factor for overall survival (OS), progression-free survival (PFS) and time to transformation (TTT) in patients with subsequently transformed FL. Remnants of FDC meshworks at transformation were associated with shorter OS and PFS from transformation. High degrees of intrafollicular CD68+ and PD-L1+ macrophage infiltration, high total area scores and an extrafollicular/diffuse pattern of FOXP3+ T cells and high intrafollicular scores of CD4+ T cells at diagnosis were associated with shorter TTT. Scores of several T-cell subset markers from the combined patient cohorts were predictive for transformation, especially CD4 and CD57.