Project description:Primary outcome(s): Measure auto-lysosome function in the cytoplasm by electron microscopy to examine the changes in the number of autophagy.
| 2626385 | ecrin-mdr-crc
Project description:Real-time metagenomics-based diagnosis of community-acquired meningitis: a prospective series, Southern France.
Project description:Endometriosis (EM) is a well-recognized disease that causes infertility in women of reproductive age. Tubal EM adversely affects fallopian tube function and female reproductive outcomes. Our study aimed to evaluate the effect of tubal EM on tubal epithelial ultrastructure and to explore the possible mechanisms of tubal infertility by analyzing the differential expression of exosomal microRNAs (miRNAs) in tubal fluid. We obtained human fallopian tube epithelium and tubal fluid samples from patients with and without tubal EM. We used scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to assess ultrastructural changes. Exosomal miRNAs in tubal fluid were extracted for microarray. We visualized epithelial damage in the tubal EM group using electron microscopy. The number of organelles decreased, and organelle structure was destroyed. A total of 14 differentially expressed exosomal miRNAs were detected in tubal fluid (fold change >2 and P value <0.05). Four miRNAs (miR-1273f, miR-5699-5p, miR-6087, miR-6747-5p) were validated by quantitative real-time polymerase chain reaction (qRT-PCR). Bioinformatic analysis showed that most of the target genes participated in embryo transport, regulation of cell communication, anatomical structure morphogenesis, and immune system processes. We verified that tubal EM damaged the tubal epithelial ultrastructure in human specimens and identified differentially expressed exosomal miRNAs in tubal liquid. These findings may elucidate the pathogenesis of tubal EM-associated infertility.
Project description:Human induced pluripotent stem cells (hiPSCs) have been used extensively in vitro to model early events in neurodevelopment. Because of a number of shortcomings, previous work has established a potential to use these cells in vivo after transplantation into the mouse brain. Here, we describe a systematic approach for the analysis of transplanted hiPSC-derived neurons and glial cells over time in the mouse brain. Using functional two-photon imaging of GCaMP6fexpressing human neural cells, we define and quantify the embryonic-like features of their spontaneous activity. This is substantiated by detailed electron microscopy (EM) of the graft. We relate this to the synaptic development the neurons undergo up to seven months in vivo. This system can now be used further for the genetic or experimental manipulation of developing hiPSC-derived cells addressing neurodevelopmental diseases like schizophrenia or Autism Spectrum Disorder.
Project description:Purpose: We aimed to investigate the effect of several anti-leukemia drugs in combination with decitabine (DAC) on the proliferation of myeloid leukemia cells in vitro and in vivo, to select the most efficient combination group and explore associated mechanisms of these combination therapies. Experimental Design: After comparing with five anti-leukemia drugs in several different kinds of cell lines, the combination effect of idarubicin (IDA) with DAC was best. In vivo, by using microPET, TUNEL, and transmission electron microscopy, the inhibitory effects obtained by sequentially combining DAC with IDA, evidenced by evaluating tumor cell proliferation and cell apoptosis. Molecular studies were conducted using gene chip, which was used to explore associated pathways, and real-time quantitative reverse transcription-PCR, western blot and immunohistochemistry (IHC), used to assess regulation of Wnt/β-catenin pathway. Results: The sequential combination of DAC and IDA showed synergistic induction of cell death in U937, HEL, SKM-1 and cells isolated from AML patients. Importantly, the inhibition of tumor growth in the sequential combination group was found to be significantly higher than that of single drug group or control group in vivo. Moreover, sequential treatment with DAC and IDA induced apoptosis and depression of the Wnt/β-catenin pathway in both culture and animal studies. Conclusions: Our findings showed that sequentially combining decitabine with idarubicin had a synergistic anti-leukemia effect. These findings were attributed to demethylation of Wnt pathway inhibitors and downregulation of Wnt pathway nuclear targets observed in vitro and in vivo. After comparing with five anti-leukemia drugs in several different kinds of cell lines, the combination effect of idarubicin (IDA) with DAC was best. In vivo, by using microPET, TUNEL, and transmission electron microscopy, the inhibitory effects obtained by sequentially combining DAC with IDA, evidenced by evaluating tumor cell proliferation and cell apoptosis. Molecular studies were conducted using gene chip, which was used to explore associated pathways, and real-time quantitative reverse transcription-PCR, western blot and immunohistochemistry (IHC), used to assess regulation of Wnt/β-catenin pathway.
