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Chronic kidney disease (CKD) is a common chronic disease that seriously threatens human life and health. Increasing evidence shows that long noncoding RNAs (lncRNA) were associated with cancer-related diseases and could function as a competing endogenous RNA (ceRNA). However, the roles and functions of the lncRNA-miRNA-mRNA network in CKD are still unclear. In this study, we performed differential expression analysis of lncRNAs, miRNAs and mRNAs in CKD using the datasets GSE66494 and GSE80247 from Gene Expression Omnibus (GEO) database. A total of 33 lncRNAs, 20 miRNAs and 240 mRNAs were differentially expressed between CKD and healthy control. Two ceRNA interaction modules composed of 11 hub nodes including 2 lncRNAs (LINC01086, LINC01094), 2 miRNAs (hsa-miR-197-3p, hsa-miR-513b-5p) and 7 mRNAs (CENPF, TOP2A, ARHGAP11A, CEP55, MELK, DTL, ANLN) were constructed according to bioinformatics analysis protocols. In vitro, knockdown of LINC01094 expression in renal tubular epithelial HK2 cells can significantly reduce the phenotype of TGFβ1-induced cell fibrosis. RNA immunoprecipitation (RIP) experiment and Dual-luciferase reporter experiments based on mutant construction validated that LINC01094 could mediate MELK expression through sponging miR-513b-5p. Our observations indicate that lowering expression of LINC01094 can significantly reduce the TGFβ1-induced HK2 cell fibrosis phenotype and inhibit the progression of renal inflammation through miR-513b-5p/MELK/smad3 signaling pathway.

MINI: We identified differentially expressed genes (DEGs) that may be involved in the development of neurofibromatosis type I by whole-transcriptional sequencing. Seven hundred eighty DEGs were identified which include protein coding genes, miRNAs, and lncRNAs. The enrichment analysis may reveal pathways that these DEGs involved. A total of 383 protein-pairs for DEGs may unfold the possible mechanism how the disease is developed.This is a clinical basic study on neurofibromatosis type I (NF-1) with spinal deformity.The current research focuses on screening key molecules affecting NF-1 with spinal deformity by transcriptome sequencing and discovering its underlying molecular biological mechanisms.NF-1 is a complex multisystem human disorder, which is often found in spinal deformities patients. The success rate of orthopedic surgery for neurofibromatosis type I combined with spinal deformities patients was low because of the lack of molecular pathology.In our study, the transcriptome-wide sequencing was preformed to identify the differentially expressed genes (DEGs) involved in this disease.Seven hundred eighty DEGs were identified which include protein coding genes, miRNAs, and lncRNAs. The DO, GO, KEGG and Reactome enrichment analysis may reveal pathways that these DEGs involved. And the 383 protein-pairs for DEGs that are involved in NF-1 combined with spinal deformities may unfold the possible mechanism how this disease is developed.The differentially expressed miRNAs and lncRNAs may contribute the ceRNA network. We focused on three key DEGs: FGFR2, MAP3K1 and STAT4. FGFR2 and MAP3K1 are members of the RAS/RAF/MEK/ERK-signaling pathway, and STAT4 were involved in the JAK/STAT pathway. The expression changes were verified by other researches and the functional cross-talk between the Ras/MAPK and JAK/STAT pathways may contribute in the disease development. This study took insight of the molecular mechanism of this disease. More detailed interactions between these factors are needed to be further explored. These key DEGs and involved pathways may provide clues in the clinical process for patients with NF-1, especially in prognosis prediction.N/A.This is a clinical basic study on neurofibromatosis type I (NF-1) with spinal deformity. The current research focuses on screening key molecules affecting NF-1 with spinal deformity by transcriptome sequencing and discovering its underlying molecular biological mechanisms. NF-1 is a complex multisystem human disorder, which is often found in spinal deformities patients. The success rate of orthopedic surgery for neurofibromatosis type I combined with spinal deformities patients was low because of the lack of molecular pathology. In our study, the transcriptome-wide sequencing was preformed to identify the differentially expressed genes (DEGs) involved in this disease. Seven hundred eighty DEGs were identified which include protein coding genes, miRNAs, and lncRNAs. The DO, GO, KEGG and Reactome enrichment analysis may reveal pathways that these DEGs involved. And the 383 protein-pairs for DEGs that are involved in NF-1 combined with spinal deformities may unfold the possible mechanism how this disease is developed. The differentially expressed miRNAs and lncRNAs may contribute the ceRNA network. We focused on three key DEGs: FGFR2, MAP3K1 and STAT4. FGFR2 and MAP3K1 are members of the RAS/RAF/MEK/ERK-signaling pathway, and STAT4 were involved in the JAK/STAT pathway. The expression changes were verified by other researches and the functional cross-talk between the Ras/MAPK and JAK/STAT pathways may contribute in the disease development. This study took insight of the molecular mechanism of this disease. More detailed interactions between these factors are needed to be further explored. These key DEGs and involved pathways may provide clues in the clinical process for patients with NF-1, especially in prognosis prediction. Level of Evidence: N/A.

The aim of the present study was to explore the expression changes of P2Y purinergic receptor 1 (P2Y1) in the distal colonic submucosa of opioid-induced constipation (OIC) rats and its association with the occurrence of OIC, an OIC rat model was generated by intraperitoneal injection of loperamide hydrochloride, a selective agonist of µ-opioid receptors (MORs). At 7 days post-treatment, the model was assessed by analyzing stool scores and calculating the gastrointestinal (GI) transit ratio of rats. The distribution of P2Y1-expressing neurons in the colonic submucosal plexus was demonstrated by immunofluorescence (IF). Western blotting was performed to evaluate the expression changes of MOR, P2Y1 and ATP synthase subunit β (ATPB) proteins in the colonic submucosa, while reverse transcription-quantitative PCR (RT-qPCR) analysis was performed to determine the relative mRNA expression of MOR and P2Y1. After 7 days, the feces of OIC rats exhibited an appearance of sausage-shaped pieces and both the stool weight and GI transit ratio of OIC rats were significantly decreased. IF revealed co-expression of P2Y1 and calbindin and MOR and ATPB in the nerve cells of the distal colonic submucosal plexus. Moreover, RT-qPCR analysis showed that the MOR mRNA levels were significantly increased in the distal colonic submucosa of OIC rats, while mRNA levels of P2Y1 were decreased. WB showed that in the distal colonic submucosa of OIC rats, MOR protein expression was increased, whereas that of P2Y1 was significantly decreased. GI transit ratio analysis suggested that the P2Y agonist ATP significantly relieved constipation symptoms in rats, while the P2Y inhibitor MRS2179 aggravated these symptoms. Finally, P2Y1 expression change was shown to be associated with the occurrence of OIC, while expression of MOR and P2Y1 was associated with OIC development in rats.

Aims: To explore the expression changes of P2Y1 in the distal colonic submucosa of opioid induced constipation (OIC) rats and its correlation with the occurrence of OIC. Methods: OIC model was generated by intraperitoneal injection of loperamide hydrochloride, a selective agonist of the μ-opioid receptor (MOR). Seven days later, the model was assessing by detecting the fecal traits and calculating the fecal water cotent. The distribution of MOR-containing neurons and P2Y1-containing neurons in colonic submucosal plexus of rat were demonstrated by immunofluorescence histochemistry. Western Blot was used to evaluate the expression changes of MOR, P2Y1 and ATP synthase subunit beta (ATPB) in colonic submucosa, while the RT-PCR analysis was performed to determine the relative mRNA expression of MOR, P2Y1 and ATPB. Results: After seven days, the feces of OIC rats had an appearance of like sausage-shaped pieces, and the fecal water content, stool weight of OIC rats were decreased. Immunofluorescence histochemistry showed the co-expression of MOR and ATPB, P2Y1 and calbindin (CB) in the nerve cells of distal colonic submucosal plexus. RT-PCR showed that MOR mRNA levels were significantly increased in the distal colonic submucosa of OIC rats, while the mRNA levels of P2Y1 were decreased. Western blot results showed that MOR protein expression was increased, and the P2Y1 protein expression was significantly decreased in the distal colonic submucosa of OIC rats.Conclusion: P2Y1 is associated with the occurrence of OIC in rats, and the expression of MOR and P2Y1 and OIC are correlated with each other.

Immune-complex glomerulonephritis (ICGN) is a major cause of nephrotic syndrome in adults and children. Cationic BSA (c-BSA) intravenous injection could produce significant albuminuria within a short time, and is a suitable in vivo experimental animal model to investigate the pathophysiology of ICGN and for drug screening, but lack of thorough study to clarify its dynamic pathophysiological alteration so far, as well as detailed changes in mRNA and LncRNA levels. The purpose of this study is to investigate the dynamic alteration in renal function, lipid metabolism and histopathology during the progress of c-BSA induced ICGN. RNA sequencing was used to identified differentially expressed mRNA and LncRNA in kidney cortex of ICGN. Results demonstrated that c-BSA induced ICGN model could completely exhibit clinical features of immune-mediated nephrotic syndrome with gradual declining renal function, and increased albuminuria and deteriorated histopathological injuries. The correlation analysis suggested that complement activation was the most key element in mediating of ICGN. RNA sequencing using rat kidney tissues combined with Gene Expression Omnibus (GEO) data of human glomerulonephritis showed the most enriched KEGG pathways in ICGN were Toll-like receptor signaling pathway, B cell receptor and Focal adhesion. The differential lncRNAs in ICGN rats were also screened, and the lncRNA-mRNA co-expression network was constructed to clarify lncRNA role in molecular mechanism of ICGN progression. Their human homogenous lncRNAs were also identified, such as ST3GAL5-AS1 and DIO3OS, which provide the potential lncRNA targets to treat ICGN. All the differential LncRNAs in ICGN kidneys caused by MMF were also identified and provided another possible pharmacological mechanism of MMF through lncRNA regulation. In summary, the current study firstly described the dynamic physiological changes of c-BSA induced ICGN, identified most key KEGG pathways, and provided lncRNA-mRNA regulatory network in ICGN.

Background Though considerable efforts have been made to improve the treatment of epithelial ovarian cancer (EOC), the prognosis of patients has remained poor. Identifying differentially expressed genes (DEGs) involved in EOC progression and exploiting them as novel biomarkers or therapeutic targets is of great value. Methods Overlapping DEGs were screened out from three independent gene expression omnibus (GEO) datasets and were subjected to Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analyses. The protein-protein interactions (PPI) network of DEGs was constructed based on the STRING database. The expression of hub genes was validated in GEPIA and GEO. The relationship of hub genes expression with tumor stage and overall survival and progression-free survival of EOC patients was investigated using the cancer genome atlas data. Results A total of 306 DEGs were identified, including 265 up-regulated and 41 down-regulated. Through PPI network analysis, the top 20 genes were screened out, among which 4 hub genes, which were not researched in depth so far, were selected after literature retrieval, including CDC45, CDCA5, KIF4A, ESPL1. The four genes were up-regulated in EOC tissues compared with normal tissues, but their expression decreased gradually with the continuous progression of EOC. Survival curves illustrated that patients with a lower level of CDCA5 and ESPL1 had better overall survival and progression-free survival statistically. Conclusion Two hub genes, CDCA5 and ESPL1, identified as probably playing tumor-promotive roles, have great potential to be utilized as novel therapeutic targets for EOC treatment.

Background: Scoliosis is a common manifestation of neurofibromatosis type 1, causing significant morbidity. The etiology of dystrophic scoliosis in neurofibromatosis type 1(NF1) is not fully understood and its therapies are lacking. This article focused on how GPR56 affected the development of NF1 cells and the related factors of osteoblast and osteoclast. Methods: Through RNA-sequencing, G-protein-coupled receptor (GPR56) was found highly differential expressed in the NF1 with scoliosis samples. We measured the GPR56 how affected the NF1 cells in vitro. Then we tested the influence of GPR56 in osteoclasts and osteoblasts. Results: We reported that knockdown GPR56 promoted the proliferation and cell cycle progression of NF1 cell. Furthermore, knockdown GPR56 can inhibited the osteoblastic differentiation and osteoclast formation, but had greater influence on osteoblastic differentiation, which indicated that knockdown GPR56 can promote the deterioration of NF1 tumors, meanwhile break the dynamic balance of bone formation and bone absorption, leading to increased bone resorption and eventually scoliosis. Conclusions: In conclusion, we found that GPR56 may inhibit the proliferation and cell cycle progression of NF1. Furthermore, GPR56 can promote osteoblastic differentiation and osteoclast formation, but had greater influence on osteoblastic differentiation, which indicated that knockdown GPR56 can promote the deterioration of NF1 tumors, meanwhile break the dynamic balance of bone formation and bone absorption, leading to increased bone resorption.

Background: Adrenocortical carcinoma (ACC) is a heterogeneous and rare malignant tumor associated with a poor prognosis. The molecular mechanisms of ACC remain elusive and more accurate biomarkers for the prediction of prognosis are needed.Methods: In this study, integrative profiling analyses were performed to identify novel hub genes in ACC to provide promising targets for future investigation. Three gene expression profiling datasets in the GEO database were used for the identification of overlapped differentially expressed genes (DEGs) following the criteria of adj.P.Value0.5 in ACC. Novel hub genes were screened out following a series of processes: the retrieval of DEGs with no known associations with ACC on Pubmed, then the cross-validation of expression values and significant associations with overall survival in the GEPIA2 and starBase databases, and finally the prediction of gene-tumor association in the GeneCards database.Results: Four novel hub genes were identified and two of them, TPX2 and RACGAP1, were positively correlated with the staging. Interestingly, co-expression analysis revealed that the association between TPX2 and RACGAP1 was the strongest and that the expression of HOXA5 was almost completely independent of that of RACGAP1 and TPX2. Furthermore, the PPI network consisting of four novel genes and seed genes in ACC revealed that HOXA5, TPX2, and RACGAP1 were all associated with TP53. Conclusions: This study identified four novel hub genes (TPX2, RACHAP1, HXOA5 and FMO2) that may play crucial roles in the tumorigenesis and the prediction of prognosis of ACC.