ABSTRACT
Objective To discover critical genes contributing to the stemness and maintenance of spermatogonial stem cells (SSCs) and provide new insights into the function of the leucine-rich repeat (LRR) family member Lrrc34 (leucine-rich repeat-containing 34) in SSCs from mice. Methods Bioinformatic methods, including differentially expressed gene (DEG), gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, were used to uncover latent pluripotency-related genes. Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence analyses were utilized to verify the mRNA and protein expression levels, respectively. RNA interference of Lrrc34 using siRNA was performed to detect its transient impact on SSCs. Results Eight DEGs between ID4-EGFP+ (G) and ID4-EGFP+/TSPAN8High (TH), eight DEGs between G and ID4-EGFP+/TSPAN8Low (TL) and eleven DEGs between TH and TL were discovered, and eleven protein-protein interaction (PPI) modules were found to be significant in the PPI network of DEGs. One of the DEGs, Lrrc34, was selected as a potential pluripotency-related gene due to its differential expression among ID4-EGFP+ spermatogonia subsets and its interaction with fibroblast growth factor 2 in the fifth module. Immunofluorescence experiments exhibited specific expression of Lrrc34 in a subpopulation of undifferentiated spermatogonia marked by LIN28A, and RT-PCR experiments confirmed the high expression of Lrrc34 in SSCs from P7 and adult mice. The transient knockdown of Lrrc34 in SSCs resulted in reduced colony sizes and significant changes in the transcriptome and apoptotic pathways. Conclusion Lrrc34 is highly expressed in mouse SSCs and is required for SSC proliferation in vitro through effects on transcriptome and signaling transduction pathways.
Subject(s)
Animals , Humans , Male , Apoptosis/genetics , Cell Proliferation/genetics , Cells, Cultured , Gene Expression Profiling/methods , Gene Ontology , Gene Regulatory Networks , Mice, Inbred C57BL , Mice, Transgenic , RNA Interference , Repressor Proteins/metabolism , Signal Transduction/genetics , Stem Cells/metabolismABSTRACT
Objective To explore the clinical effect of ulinastatin combined with recruitment maneuver on extrapulmonary acute respiratory distress syndrome (ARDS). Methods Forty-two patients with extrapulmonary ARDS, who received ulinastatin combined with recruitment maneuver in Eastern Hepatobiliary Surgery Hospital of Second Military Medical University from Jun. 2014 to Jun. 2016, were assigned to experimental group; and 45 patients with extrapulmonary ARDS, who were treated by recruitment maneuver without ulinastatin at the same time, were taken as control group. The blood gas analyzer was used to record radial arterial oxygen partial pressure (PaCO2), carbon dioxide partial pressure (PaCO2) and oxygenation index (OI) before and after treatment in the two groups; the breathing machine was used to record the inspiratory peak pressure (PIP), plateau pressure (Pplat), static lung compliance (Cs) and dynamic lung compliance (Cd); and ELISA was used to detect the serum levels of IL-6, TNF-α and IL-10. Results PaCO2 on 2nd, 3rd, 5th, 6th and 7th day after treatment, PaCO2 on 1st, 3rd, 5th and 7th day after treatment, and OI on 6th and 7th day after treatment in the experimental group were higher than those in the control group (all P<0.05). After treatment, PIP and Pplat in the two groups were significantly decreased (P<0.05), and Cs and Cd were significantly increased (P<0.05); the changes in the experimental group were significantly greater compared with the control group (P<0.05). In the experimental group and the control group, IL-6 and TNF-α levels were significantly decreased (P<0.05), and IL-10 levels were increased (P<0.05) after treatment; the changes in the experimental group were significantly greater compared with the control group (P<0.05). Conclusion Ulinastatin combined with recruitment maneuver can more effectively reduce lung injury and improve pulmonary ventilation of the patients with extrapulmonary ARDS compared with simple recruitment maneuver.
ABSTRACT
The role of ROS in hydroquinone-induced inhibition of K562 cell erythroid differentiation was investigated. After K562 cells were treated with hydroquinone for 24 h, and hemin was later added to induce erythroid differentiation for 48 h, hydroquinone inhibited hemin-induced hemoglobin synthesis and mRNA expression of γ-globin in K562 cells in a concentration-dependent manner. The 24-h exposure to hydroquinone also caused a concentration-dependent increase at an intracellular ROS level, while the presence of N- acetyl-L-cysteine prevented hydroquinone- induced ROS production in K562 cells. The presence of N-acetyl-L-cysteine also prevented hydroquinone inhibiting hemin-induced hemoglobin synthesis and mRNA expression of γ-globin in K562 cells. These evidences indicated that ROS production played a role in hydroquinone-induced inhibition of erythroid differentiation.