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2022 ◽  
Author(s):  
Liming Jin ◽  
Zhaoxia Zhang ◽  
Zhang Wang ◽  
Xiaojun Tan ◽  
Zhaoying Wang ◽  
...  

Abstract Background: CSCs play an important role in tumor development. Some studies have demonstrated that piRNAs participate in the progression of various cancers. However, the detailed function of piRNAs in CSCs requires further investigation. This study aimed to investigate the significance of some piRNAs in Piwil2-iCSCs. Methods and Results: Differentially expressed piRNAs in Piwil2-iCSCs were screened by high-throughput sequencing. Target genes were predicted by the miRanda algorithm and subjected to GO and KEGG analysis. One of the differential piRNAs, novel piRNA MW557525, was transfected and its target gene NOP56 was silenced in Piwil2-iCSCs, respectively. RT-qPCR, western blot and dual luciferase reporter assay were used to investigate the interaction of piRNA MW557525 and NOP56. We identified the effect of piRNA MW557525 and NOP56 knockdown on cell proliferation, migration, invasion, and apoptosis via CCK-8, transwell assay, and flow cytometry. The expressions of CD24, CD133, KLF4, and SOX2 were detected via WB. The results showed that piRNA MW557525 was negatively correlated with NOP56, and it promoted the proliferation, migration, invasion, and inhibited apoptosis in Piwil2-iCSCs, and it also promoted the expressions of CD24, CD133, KLF4, and SOX2, while NOP56 showed the opposite effect. Conclusions: These findings suggested that novel piRNA MW557525 might be a novel therapeutic target in Piwil2-iCSCs.


2022 ◽  
Author(s):  
Xu-Peng Wen ◽  
Guo Long ◽  
Yue-Zhong Zhang ◽  
He Huang ◽  
Tao-Hua Liu ◽  
...  

Abstract Background: Acute respiratory distress syndrome (ARDS) is characterized by refractory hypoxemia caused by accumulation of pulmonary fluid, which is related to inflammatory cell infiltration, impaired tight junction of pulmonary epithelium and impaired Na, K-ATPase function, especially Na, K-ATPase α1 subunit. Up until now, the pathogenic mechanism at the level of protein during lipopolysaccharide- (LPS-) induced ARDS remains unclear.Methods: Using an unbiased, discovery and quantitative proteomic approach, we discovered the differentially expressed proteins binding to Na, K-ATPase α1 between LPS-A549 cells and Control-A549 cells. These Na, K-ATPase α1 interacting proteins were screened by co-immunoprecipitation (Co-IP) technology. Among them, some of the differentially expressed proteins with significant performance were identified and quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The protein interaction network was constructed by the related Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Several differentially expressed proteins were validated by Western blot.Results: Of identified 1598 proteins, 89 were differentially expressed proteins between LPS-A549 cells and Control-A549 cells. Intriguingly, protein-protein interaction network showed that there were 244 significantly enriched co-expression among 60 proteins in the group control-A549. while the group LPS-A549 showed 43 significant enriched interactions among 29 proteins. The related GO and KEGG analysis found evident phenomena of ubiquitination and deubiquitination, as well as the pathways related to autophagy. Among proteins with rich abundance, there were several intriguing ones, including the deubiquitinase (OTUB1), the tight junction protein zonula occludens-1 (ZO-1), the scaffold protein in CUL4B-RING ubiquitin ligase (CRL4B) complexes (CUL4B) and the autophagy-related protein sequestosome-1 (SQSTM1).Conclusions: In conclusion, our proteomic approach revealed targets related to the occurrence and development of ARDS, being the first study to investigate significant differences in Na, K-ATPase α1 interacting proteins between LPS-induced ARDS cell model and control-A549 cell. These proteins may help the clinical diagnosis and facilitate the personalized treatment of ARDS.


2022 ◽  
Vol 22 (1) ◽  
Author(s):  
Jing Xia ◽  
Mengjie Wang ◽  
Yi Zhu ◽  
Chaozhi Bu ◽  
Tianyu Li

Abstract Background Long non-coding RNAs (lncRNAs) are transcripts longer than 200 nucleotides (nt) that are involved in the pathogenesis and development of various cancers including B cell acute lymphoblastic leukemia (B–ALL). To determine the potential roles of lncRNAs involved in pathogenesis of B-ALL, we analyzed the expression profile of lncRNAs and mRNAs in B-ALL, respectively, and constructed lncRNAs/mRNAs interaction network. Methods We performed RNA sequencing of 10 non-leukemic blood disease donors and 10 B-ALL patients for Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Interactions among mRNAs were predicted using the STRING database. Quantitative real time PCR (qRT-PCR) was performed to verify the RNA-seq data of lncRNAs and mRNAs. Potential functions of subtype-specific lncRNAs were determined by using coexpression-based analysis on distally (trans-pattern) located protein-coding genes. Results A total of 1813 differentially expressed transcripts (DETs) and 2203 lncRNAs were identified. Moreover, 10 dysregulated lncRNAs and 10 mRNAs were randomly selected, and further assessed by RT-qPCR in vitro. Go and KEGG analysis demonstrated that the differentially expressed mRNAs were most closely associated with myeloid leukocyte activation and in transcriptional misregulation in cancer, respectively. In addition, co-expression analysis demonstrated that these lncRNAs, including MSTRG.27994.3, MSTRG.21740.1, ENST00000456341, MSTRG.14224.1 and MSTRG.20153.1, may mediate the pathogenesis and development of B-ALL via lncRNA-mRNA network interactions. Conclusions These results showed that several mRNAs and lncRNAs are aberrantly expressed in the bone marrow of B-ALL patients and play potential roles in B-ALL development, and be useful for diagnostic and/or prognostic purposes in pediatric B–ALL. Data availability The datasets used during our study are available through HARVARD Dataverse Persistent ID doi:10.7910/DVN/LK9T4Z.


2022 ◽  
Author(s):  
Biyu Shen ◽  
Songsong Shi ◽  
Haoyang Chen ◽  
Yi Lu ◽  
Hengmei Cui ◽  
...  

Abstract Background and Objective: Fanconi anemia (FA) patients have a reduced ability to form blood cells, accompanied by multiple congenital malformations, mental retardation, solid tumors, and other symptoms. However, the molecular mechanism that causes FA is unclear, and few studies have addressed the regulatory mechanism of immune infiltration in FA. Here, we aimed to identify differentially expressed genes (DEGs), pathways, and immune infiltration involved in FA using integrated bioinformatics analysis and molecular mechanisms. Methods: The GEO gene chip database was searched for FA low density bone marrow tissue, and the content and proportion of 22 types of immune cells in the FA group and the normal group were analyzed using CIBERSORT. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of FA differentially expressed genes (DEGs) using R language and related package programs was also performed.Results: The expression levels of T cells regulatory (Tregs), M2 macrophages, T cells CD8, dendritic cells resting, and T cells CD4 naïve in FA were higher than in the normal group. Furthermore, the expression levels of naïve B cells, monocytes, and resting mast cells in FA were lower than in the normal group. GO analysis of FA differential genes showed that “neutrophil degranulation,” “neutrophil activation,” and “neutrophil activation involved in immune response,” were most frequently enriched among biological processes, with “specific granule,” “tertiary granule,” “tertiary granule lumen” among cellular components, and “carbohydrate binding” among molecular functions. For the KEGG analysis, “Asthma” was most often enriched.Conclusion: This study obtained useful data related to immune infiltration, DEGs, and gene pathways of FA, and provides new evidence for immunotherapy and clinical assessment of FA patients. These results are potentially a useful reference for subsequent related scientific research.


Genes ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 32
Author(s):  
Xiaolan Zhang ◽  
Pengjia Bao ◽  
Na Ye ◽  
Xuelan Zhou ◽  
Yongfeng Zhang ◽  
...  

The development of hair follicles in yak shows significant seasonal cycles. In our previous research, transcriptome data including mRNAs and lncRNAs in five stages during the yak hair follicles (HFs) cycle were detected, but their regulation network and the hub genes in different periods are yet to be explored. This study aimed to screen and identify the hub genes during yak HFs cycle by constructing a mRNA-lncRNA co-expression network. A total of 5000 differently expressed mRNA (DEMs) and 729 differently expressed long noncoding RNA (DELs) were used to construct the co-expression network, based on weighted genes co-expression network analysis (WGCNA). Four temporally specific modules were considered to be significantly associated with the HFs cycle of yak. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that the modules are enriched into Wnt, EMC-receptor interaction, PI3K-Akt, focal adhesion pathways, and so on. The hub genes, such as FER, ELMO1, PCOLCE, and HOXC13, were screened in different modules. Five hub genes (WNT5A, HOXC13, DLX3, FOXN1, and OVOL1) and part of key lncRNAs were identified for specific expression in skin tissue. Furthermore, immunofluorescence staining and Western blotting results showed that the expression location and abundance of DLX3 and OVOL1 are changed following the process of the HFs cycle, which further demonstrated that these two hub genes may play important roles in HFs development.


Animals ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 16
Author(s):  
Peng Ding ◽  
Huichao Liu ◽  
Yueyue Tong ◽  
Xi He ◽  
Xin Yin ◽  
...  

Although the fertilized eggs were found to contain microbes in early studies, the detailed composition of yolk microbiota and its influence on embryo intestinal microbiota have not been satisfactorily examined yet. In this study, the yolk microbiota was explored by using 16s rRNA sequencing at different developmental stages of the broiler embryo. The results showed that the relative abundance of yolk microbiota was barely changed during embryogenesis. According to the KEGG analysis, the yolk microbiota were functionally related to amino acid, carbohydrate, and lipid metabolisms during chicken embryogenesis. The yolk microbiota influences the embryonic intestinal microbiota through increasing the colonization of Proteobacteria, Firmicutes, and Bacteroidetes in the intestine, particularly. The intestinal microbes of neonatal chicks showed higher proportions of Faecalibacterium, Blautia, Coprococcus, Dorea, and Roseburia compared to the embryonic intestinal microbiota. Our findings might give a better understanding of the composition and developmental change of yolk microbiota and its roles in shaping the intestinal microbiota.


Horticulturae ◽  
2021 ◽  
Vol 7 (12) ◽  
pp. 580
Author(s):  
Zhixing Nie ◽  
Jianying Chen ◽  
Yunpeng Song ◽  
Hongfei Fu ◽  
Hong Wang ◽  
...  

Cytoplasmic male-sterility (CMS) is important for the utilization of crop heterosis and study of the molecular mechanisms involved in CMS could improve breeding programs. In the present study, anthers of the pepper CMS line HZ1A and its maintainer line HZ1B were collected from stages S1, S2, and S3 for transcriptome sequencing. A total of 47.95 million clean reads were obtained, and the reads were assembled into 31,603 unigenes. We obtained 42 (27 up-regulated and 15 down-regulated), 691 (346 up-regulated and 345 down-regulated), and 709 (281 up-regulated and 428 down-regulated) differentially expressed genes (DEGs) in stages S1, S2, and S3, respectively. Through Gene Ontology (GO) analysis, the DEGs were found to be composed of 46 functional groups. Two GO terms involved in photosynthesis, photosynthesis (GO:0015986) and photosystem I (GO:0009522), may be related to CMS. Through Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, oxidative phosphorylation (ko00190) and phenylpropanoid biosynthesis (ko00940) were significantly enriched in the S1 and S2 stages, respectively. Through the analysis of 104 lipid metabolism-related DEGs, four significantly enriched KEGG pathways may help to regulate male sterility during anther development. The mitochondrial genes orf470 and atp6 were identified as candidate genes of male sterility for the CMS line HZ1A. Overall, the results will provide insights into the molecular mechanisms of pepper CMS.


Author(s):  
Nan Xiong ◽  
Qiangming Sun

At present, there are still no specific therapeutic drugs and appropriate vaccines for Dengue. Therefore, it is very important to explore distinct clinical diagnostic indicators. In this study, we combined differentially expressed genes (DEGs) analysis and weighted co-expression network analysis (WGCNA) to screen a stable and robust biomarker which can be used to distinguish three clinical stages of Dengue and severity of Dengue. CD38 can distinguish excellently Early Acute, Late Acute, Convalescent stages for Dengue patients, and ZNF595 can discriminate DHF from DF in whole acute stages. We also found that three clinical stages can be discriminated based on the fractions of Plasma cells, activated memory CD4+ T cells, and Monocytes. In different clinical stages different immune cells function positively. Negative inhibition of viral replication based on Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and Gene set enrichment analysis (GSEA), up-regulated autophagy genes and impairing immune system are potential reasons resulting in dengue hemorrhagic fever (DHF).


2021 ◽  
Vol 2021 ◽  
pp. 1-10
Author(s):  
Yi Xie ◽  
Kainan Zhou ◽  
Yan Wang ◽  
Shuhan Yang ◽  
Suying Liu ◽  
...  

Background. Cancer-related fatigue (CRF) is an increasingly appreciated complication in cancer patients, which severely impairs their quality of life for a long time. Astragali Radix (AR) is a safe and effective treatment to improve CRF, but the related mechanistic studies are still limited. Objective. To systematically analyze the mechanism of AR against CRF by network pharmacology. Methods. TCMSP was searched to obtain the active compounds and targets of AR. The active compound-target (AC-T) network was established and exhibited by related visualization software. The GeneCards database was searched to acquire CRF targets, and the intersection targets with AR targets were used to make the Venny diagram. The protein-protein interaction (PPI) network of intersection targets was established, and further, the therapeutic core targets were selected by topological parameters. The selected core targets were uploaded to Metascape for GO and KEGG analysis. Finally, AutoDock Vina and PyMOL were employed for molecular docking validation. Results. 16 active compounds of AR were obtained, such as quercetin, kaempferol, 7-O-methylisomucronulatol, formononetin, and isorhamnetin. 57 core targets were screened, such as AKT1, TP53, VEGFA, IL-6, and CASP3. KEGG analysis manifested that the core targets acted on various pathways, including 137 pathways such as TNF, IL-17, and the AGE-RAGE signaling pathway. Molecular docking demonstrated that active compounds docked well with the core targets. Conclusion. The mechanism of AR in treating CRF involves multiple targets and multiple pathways. The present study laid a theoretical foundation for the subsequent research and clinical application of AR and its extracts against CRF.


2021 ◽  
Author(s):  
Xialin Cheng ◽  
Tao Dai ◽  
Wu Bao ◽  
Lingxi Chen ◽  
Zexin Zhang ◽  
...  

Abstract Background Giant congenital melanocytic nevi (GCMNs) are melanotic lesion present at birth, which often cause severe psychological and financial burden to patients and their families.However, the pathogenes is still unclear. Objective We aim to identify key genes and biological processes that related to the development of GCMN. Methods We sequenced ten pairs of GCMN tissues and adjacent normal tissues by high-throughput RNA-seq, then used GO and KEGG analysis to find inportment pathways, and used MCODE,Cluego and Cytohubba plugin of Cytoscape software to identify hub genes. Results A total of 1163 differentially expressed genes were identified. 29 BPs, 18 CCs, and 17 MFs were significantly enriched in GO analysis and no pathway was significantly enriched in KEGG analysis. PPI Visual Network consisted of 779 nodes and 2359 edges,which was be divided into 25 functional modules by MCODE. We discovered most of the hub genes were located in module 5,and the top 3 hub genes (PTGS2,EGF,SOX10) in module 5 were involved in GO and KEGG enrichment pathways --“Arachidonic acid metabolism”,”glycosaminoglycan biosynthetic process”,”developmental pigmentation” respectively. Conclusion PTGS2, EGF and SOX10 are thought to be the three most important hub genes and may play essential roles in the development of GCMN.


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