Project description:The present study aimed to investigate the profile of serum-circulating miRNAs and their target genes and elucidate their role in infected and non-infected C. trachomatis RSA patients by microarray miRNAs and mRNAs were found to be differentially expressed in C. trachomatis-positive RSA patients Non-heparinized blood samples were collected from 25 RSA patients with history of three or more consecutive abortions
Project description:A total of 39 miRNAs were either up- or down- regulated by at least 1.5-fold and P value< 0.05 in the RSA, including 20 up-regulated miRNAs and 19 down-regulated miRNAs
Project description:We isolated CD14+CD45+ dMΦ from the normal or RSA decidua by flow cytometry, followed by single cell RNA sequencing (scRNA-seq). In total, 23,062 single-cell transcriptomes of macrophage were profiled (12,470 Normal and 10,592 RSA), which were divided into 13 major clusters via T-distributed stochastic neighbor embedding (t-SNE) visualization.
Project description:The cancer-associated stroma plays a critical but incompletely characterized role in melanocytic neoplasia. To define stromal transcriptional changes across the transition from normal tissue to benign nevus to malignant melanoma, laser-capture microdissection (LCM) was used to selectively isolate stromal regions from archival formalin-fixed, paraffin-embedded (FFPE) tissue sections of normal skin (n=18 patients), benign cutaneous nevi (n=10 patients), cutaneous melanoma (n=20 patients), avoiding contamination from epidermal, melanocytic, and tumour cell compartments. Where present, matched epidermal and tumour regions from the same patients were also microdissected for comparison. RNA was extracted from microdissected FFPE material and sequenced by single-end RNA sequencing (Illumina NovaSeq 6000, 100 bp). This dataset enables identification of stromal transcriptional changes conserved across the cutaneous melanocytic malignancy spectrum.
Project description:To explore the changes of local microenvironment in the feto-maternal interface during early pregnancy in RSA, we next examined the transcriptional profiles of the decidua tissue by RNA sequencing using samples from 3 RSA patients and 3 healthy controls
Project description:Recurrent spontaneous abortion (RSA) is a severe pregnancy disorder with heterogeneous etiologies. Emerging evidence has suggested a functional role of epigenetic modification in RSA development. N7-methylguanosine (m7G) in mRNA is one of the frequently reported epigenetic modifications closely associated with various human diseases. However, the role of m7G modification in the development of RSA remains poorly understood. In this study, we perform multi-omics analyses of decidua from RSA patients and healthy controls (HC), and report for the first time that m7G modification was associated with RSA development. MeRIP-seq verifies that the m7G modification features, such as sequence motifs, distribution regions, and distribution densities, were significantly different between the two groups. Notably, genes with altered m7G modification were mostly enriched in embryonic and systemic morphogenesis and development. Moreover, we found an increased expression of the m7G methyltransferase METTL1 in the RSA decidua. RNA sequencing (RNA-seq) further revealed a vastly changed transcriptome in the RSA decidua. Conjoint analysis of MeRIP-seq and RNA-seq further showed that the methylation levels and expression levels of 48 genes changed simultaneously in the RSA decidua. Furthermore, we identified 7 RSA-related genes such as SOCS3, RASA1, FLT1, and JUNB were downregulated, while GNLY was up-regulated in RSA decidua. More importantly, METTL1 showed high-affinity binding to target mRNAs harboring altered m7G modification, indicating dysregulated RSA-genes may be driven by METTL1-mediated m7G modification. These findings provide novel insight into the functional role of m7G modification in RSA pathogenesis.
Project description:Recurrent spontaneous abortion (RSA) affects 1-5% of pregnant women, yet the mechanisms behind the disease remain unknown. It is well acknowledged that insufficient blood flow and placental abnormalities are key factors contributing to RSA. The abortive mouse model was constructed by intraperitoneal injection of β2-GPI. miRNA expression profiles in placental tissues of aborted and normal pregnant mice were detected by high-throughput sequencing. We found that miR-381-3p was highly expressed in the placenta of aborted mice and may bind to VEGFA
Project description:The number of circulating tumor cells (CTCs) in metastatic prostate cancer patients provides prognostic and predictive information. However, it is the molecular characterization of CTCs that offers insight into the biology of these tumor cells in the context of personalized treatment. We performed a pilot study to evaluate the feasibility of isolation and genomic profiling of CTCs in castration-resistant prostate cancer. CTCs in 7.5 mLs of blood in 20 castration-resistant metastatic prostate cancer patients were enumerated using CellSearch. Additional 10-20 mLs of blood from 12 patients positive for CTCs were subjected to immunomagnetic enrichment and fluorescence activated cell sorting (IE/FACS) to isolate pools of ~20 CTCs. Genomic DNA of CTCs was subjected to whole genome amplification followed by gene copy number analysis via array comparative genomic hybridization (aCGH). Archival primary tumor biopsy samples available from 2 patients were also subjected to aCGH.
Project description:Recurrent spontaneous abortion (RSA), defined as the failure of two or more consecutive clinical pregnancies before 20 weeks of gestation, affects approximately 1% of couples attempting to conceive. However, the underlying mechanisms of 50% of cases are unknown. The villus plays a crucial role during pregnancy, which can provide a delicate balance between immune tolerance and defense to maintain the pregnancy.We performed RNA-seq to identify gene expression alterations in the villus of RSA patients and controls. RNA-seq was done using TruSeq Stranded mRNA Library Preparation. Briefly, intact RNA was fragmented, end repaired, adapter ligation and PCR amplified following illumina protocol. Libraries were sequencing by illumins Hiseq 2000. After quality control, sequence data were processed with STAR to generate read alignments with hg19. Raw read counts for annotated genes were obtained with featureCounts with default settings, normalized and analyzed using DEseq2.