Hsa_circRNA_102610 upregulation in Crohn’s disease promotes transforming growth factor-β1-induced epithelial-mesenchymal transition via sponging of hsa-miR-130a-3p

BACKGROUND The incidence of inflammatory bowel disease,a chronic intestinal inflammatory disorder that includes Crohn’s disease(CD)and ulcerative colitis,is rising.Circular RNAs are considered valuable diagnostic biomarkers for CD.Current evidence supports the views that epithelial-mesenchymal transition(EMT)plays an important role in CD pathogenesis,and that hsa-miR-130a-3p can inhibit transforming growth factor-β1(TGF-β1)-induced EMT.Our previous study revealed that hsa_circRNA_102610 was upregulated in CD patients.Moreover,we predicted an interaction between hsa_circRNA_102610 and hsa-miR-130a-3p.Thus,we hypothesized that hsa_circRNA_102610 may play roles in the proliferation and EMT of intestinal epithelial cells by sponging hsa-miR-130a-3p to participate in the pathogenesis of CD.AIM To explore the mechanism of hsa_circRNA_102610 in the pathogenesis of CD.METHODS The relative expression levels of hsa_circRNA_102610 and hsa-miR-130a-3p in patients were detected by quantitative reverse transcription-polymerase chain reaction.The proliferation of human intestinal epithelial cells(HIECs)and normal-derived colon mucosa cell line 460(NCM460)cells was detected by cell counting kit-8,5-ethynyl-2’-deoxyuridine staining and cell cycle assays following overexpression or downregulation of hsa_circRNA_102610.Cell proliferation assays were performed as described above in a rescue experiment with hsa-miR-130a-3p mimics.The interaction of hsa_circRNA_102610 and hsa-miR-130a-3p was verified by fluorescence in situ hybridization and dual luciferase reporter assays.The relative expression levels of CyclinD1,mothers against decapentaplegic homolog 4(SMAD4),E-cadherin,N-cadherin and Vimentin were detected by western blotting following hsa_circRNA_102610 overexpression,TGF-β1-induced EMT or hsa-miR-130a-3p mimic transfection(in rescue experiments).RESULTS Upregulation of hsa_circRNA_102610 was determined to be positively correlated with elevated fecal calprotectin levels in CD(r=0.359,P=0.007)by Pearson correlation analysis.Hsa_circRNA_102610 promoted the proliferation of HIECs and NCM460 cells,while hsa-miR-130a-3p reversed the cell proliferationpromoting effects of hsa_circRNA_102610.Fluorescence in situ hybridization and dual luciferase reporter assays showed that hsa_circRNA_102610 directly bound hsa-miR-130a-3p in NCM460 and 293T cells.An inverse correlation between downregulation of hsa-miR-130a-3p and upregulation of hsa_circRNA_102610 in CD patients was observed(r=-0.290,P=0.024)by Pearson correlation analysis.Moreover,overexpression of hsa_circRNA_102610 promoted SMAD4 and CyclinD1 protein expression validated by western-blotting.Furthermore,overexpression of hsa_circRNA_102610 promoted TGF-β1 induced EMT in HIECs and NCM460 cells via targeting of hsa-miR-130a-3p,with increased expression of Vimentin and N-cadherin and decreased expression of E-cadherin.CONCLUSION Hsa_circRNA_102610 upregulation in CD patients could promote the proliferation and EMT of intestinal epithelial cells via sponging of hsa-miR-130a-3p.

Rapid diagnosis of disseminated Mycobacterium mucogenicum infection in formalin-fixed, paraffin-embedded specimen using nextgeneration sequencing: A case report

BACKGROUND Mycobacterium mucogenicum(M.mucogenicum)belongs to the group of rapidly growing Nontuberculous mycobacteria.This microorganism is associated with a wide spectrum of infectious diseases.Due to a low detection rate or the time required for conventional culture methodology,a rapid and broad-spectrum method is necessary to identify rare pathogens.CASE SUMMARY A 12-year-old immunocompetent girl presented with painful masses for five months.The first mass was found in the right upper quadrant of the abdomen,and was about 1 cm×1.5 cm in size,tough but pliable in texture,with an irregular margin and tenderness.An abscess gradually formed and ulcerated with suppuration of the mass.Three new masses appeared on the back one by one.Chest computed tomography showed patchy and streaky cloudy opacities in both lungs.Needle aspiration of the abscess was performed,but the smear and conventional culture were negative,and the pathological examination showed no pathogens.We then performed next-generation sequencing using a formalinfixed,paraffin-embedded specimen to identify the pathogen.A significantly high abundance of M.mucogenicum was detected.The patient’s abscesses gradually decreased in size,while inflammation in both lungs improved following 12-wk of treatment.No recurrence was observed four months after the end of the one-year treatment period.CONCLUSION Next-generation sequencing is a promising tool for the rapid and accurate diagnosis of rare pathogens,even when using a formalin-fixed,paraffin-embedded specimen.

Effects of flooding on grafted annona plants of different scion/rootstock combinations

Annona atemoya Hort cv. African Pride (AP) is highly valued due to its high quality and unique flavor, but highly susceptible to water-logging. Prevalence of root diseases in saturated soils is one of the main problems in production, which restricts the development of AP in south China, where flooding frequently occurs in rainy seasons. However, some annona species, e.g. A. montana, A. glabra and A. muricata, are relatively tolerant to continuous flooding and periodic water-logging conditions, but of limited commercial value. Yet, the potential may exist to increase flood tolerance of commercial annona varieties by the use of flood tolerant rootstocks. An experiment was conducted with the aim to study the effects of continuous or periodical soil flooding on tree performances of four different annona scion/rootstock combinations: AP/AR/G (scion/interstock/rootstock), AR/G (scion/rootstock), AP/AR/M and AR/M, where AP stands for Annona atemoya Hort cv. African Pride, AR for the hybrid of “AP” atemoya × A. reticulata, used as an interstock, G for pond apple (A. glabra), and M for mountain soursop (A. montana). Plant growth, leaf net photosynthetic rates and chlorophyll fluorescence parameters were measured regularly after flooding treatments were applied. Flooding treatments reduced shoot extension, leaf production, net photosynthetic rates and maximum quantum efficiency of photosystem II (Fv/Fm) in plants of AP/AR/M and AR/M, which displayed wilting within 2 weeks of flooding, with a higher wilting percentage in AP/AR/M than in AR/M. The wilted plants shed all leaves but remained alive and sprouted new but weak shoots after 16 weeks of flooding. Long term flooding did not suppress but enhanced photosynthesis as well as tree growth in AP/AR/G and AR/G, with vigorous growth of adventitious roots. Thus, we suggest the use A. glabra instead of A. montana as a rootstock and AR as an interstock to increase flood tolerance of commercial annona varieties.