Role and Mechanism of NUP160-regulated Autophagy in Pathogenesis of Diabetic Nephropathy.

\Introduction. Diabetes mellitus (DM) is one of the most common chronic diseases worldwide, and diabetic nephropathy (DN) is the most significant complication of DM, which is highly prevalent and difficult to cure. This research project aims to investigate the role and mechanism of Nucleoporin 160kDa (NUP160)-regulated autophagy in the pathogenesis of DN. METHODS: NUP160 levels in diabetic and non-diabetic kidney tissues were measured by Western blot, and the connection between NUP160 and renal function of DN patients was analyzed. The podocytes were divided into four groups, namely the standard group (culture medium: standard glucose solution), high glucose (HG) group (HG solution), HG+si-NUP160 group (HG solution+si-NUP160 transfection) and HG+si-NC group (HG solution+si-NUP 160 transfection) for the determination of apoptosis by flow cytometry and measurements of LC3B, Prostacyclin-62 (P62), Janus kinase 2 (JAK2) and Signal transducer and activator of transcription3 (STAT3) by Western blot. RESULTS: In DN patients, NUP160 decreased in podocytes and was inversely proportional to Blood urea nitrogen (BUN), Serum creatinine (Scr) and ß2-Microglobulin (ß2-MG) (P < .05). Compared with a standard group, the apoptosis rate, P62 level, and the ratios of phosphorylation-JAK2 (p-JAK2)/JAK2, phosphorylation-STAT3 (p-STAT3)/STAT3, and LC3B-Ⅱ/LC3B-Ⅰ elevated in the other three groups (P < .05). Apoptosis rate and P62 level, p-JAK2/JAK2 and p-STAT3/STAT3 ratios increased, and LC3B-Ⅱ/LC3B-Ⅰ ratio decreased in the HG+si-NUP160 group (P < .05), while those in HG+si-NC group showed no evident changes, compared with HG group (P > .05). CONCLUSION: NUP160 is downregulated in DN and can affect cellular autophagy through the activation of JAK2/STAT3 signaling pathway.  DOI: 10.52547/ijkd.7884.

Classifying and Scoring Major Depressive Disorders by Residual Neural Networks on Specific Frequencies and Brain Regions.

Major Depressive Disorder (MDD) - can be evaluated by advanced neurocomputing and traditional machine learning techniques. This study aims to develop an automatic system based on a Brain-Computer Interface (BCI) to classify and score depressive patients by specific frequency bands and electrodes. In this study, two Residual Neural Networks (ResNets) based on electroencephalogram (EEG) monitoring are presented for classifying depression (classifier) and for scoring depressive severity (regression). Significant frequency bands and specific brain regions are selected to improve the performance of the ResNets. The algorithm, which is estimated by 10-fold cross-validation, attained an average accuracy rate ranging from 0.371 to 0.571 and achieved average Root-Mean-Square Error (RMSE) from 7.25 to 8.41. After using the beta frequency band and 16 specific EEG channels, we obtained the best-classifying accuracy at 0.871 and the smallest RMSE at 2.80. It was discovered that signals extracted from the beta band are more distinctive in depression classification, and these selected channels tend to perform better on scoring depressive severity. Our study also uncovered the different brain architectural connections by relying on phase coherence analysis. Increased delta deactivation accompanied by strong beta activation is the main feature of depression when the depression symptom is becoming more severe. We can therefore conclude that the model developed here is acceptable for classifying depression and for scoring depressive severity. Our model can offer physicians a model that consists of topological dependency, quantified semantic depressive symptoms and clinical features by using EEG signals. These selected brain regions and significant beta frequency bands can improve the performance of the BCI system for detecting depression and scoring depressive severity.

Long Noncoding RNA XIST Regulates Myocardial Infarction via miR-486-5p/SIRT1 Axis.

Myocardial infarction (MI) is severe heart disease leading to the death worldwide. Long noncoding RNAs (lncRNAs) play a vital role in progression of numerous heart diseases. In the present study, we examined the effects of lncRNA XIST and underlying mechanism on hypoxia-induced apoptosis. In vitro model of MI was established by inducing hypoxia in H9c2 cells. CCK-8 assay was used to measure the cell viability in hypoxia-induced H9c2 cells. The rate of cell apoptosis was assessed by using caspase-3 assay. Transfection was carried out to upregulate the expressions of lncRNA XIST, and miR-486-5p. RT-qPCR was used to measure the levels of lncRNA XIST and miR-486-5p. Also, the relation between XIST and miR-486-5p was confirmed by using Luciferase reporter assay. Our findings revealed that hypoxia significantly downregulated the expressions of XIST. Also, the cell viability markedly increased due to the overexpression of XIST in hypoxia-induced H9c2 cells, while overexpression of XIST significantly reduced the cell apoptosis in hypoxia-induced H9c2 cells. On the other hand, opposite effects were observed due to the downregulation of XIST in hypoxia-induced H9c2 cells. Moreover, XIST negatively regulated the expression of miR-4486-5p and upregulation of XIST inhibited hypoxic injury by downregulating miR-486-5p. Furthermore, SIRT1 expression was negatively regulated by miR-486-5p. We concluded that lncRNA XIST might provide protection against injury induced by hypoxia via miR-486-5p/SIRT1 axis.

NUP160 knockdown inhibits the progression of diabetic nephropathy in vitro and in vivo.

Diabetic nephropathy (DN) is a severe diabetic complication and podocyte damage is a hallmark of DN. The Nucleoporin 160 (NUP160) gene was demonstrated to regulate cell proliferation and apoptosis in mouse podocytes. This study explored the possible role and mechanisms of NUP160 in high glucose-triggered podocyte injury. A rat model of DN was established by intraperitoneal injection of 60 mg/kg streptozotocin (STZ). Podocytes were treated with 33 mM high glucose. The effects of the Nup160 on DN and its mechanisms were assessed using MTT, flow cytometry, Western blot, ELISA, RT-qPCR, and luciferase reporter assays. The in vivo effects of NUP160 were analyzed by HE, PAS, and MASSON staining assays. The NUP160 level was significantly upregulated in podocytes treated with 33 mM high glucose. Functionally, NUP160 knockdown alleviated high glucose-induced apoptosis and inflammation in podocytes. Mechanistically, miR-495-3p directly targeted NUP160, and lncRNA HCG18 upregulated NUP160 by sponging miR-495-3p by acting as a ceRNA. Additionally, NUP160 overexpression reversed the effects of HCG18 knockdown in high glucose treated-podocytes. The in vivo assays indicated that NUP160 knockdown alleviated the symptoms of DN rats. NUP160 knockdown plays a key role in preventing the progression of DN, suggesting that targeting NUP160 may be a potential therapeutic strategy for DN treatment.

Nucleoporin 160 (NUP160) inhibition alleviates diabetic nephropathy by activating autophagy.

Diabetic nephropathy (DN) is the leading cause of end-stage renal disease worldwide. Autophagy was reported to be related to the pathogenesis of DN. This research investigated the function of the Nucleoporin 160 (Nup160) gene in regulating autophagy in DN. A mouse model of DN was established through an intraperitoneal injection of streptozotocin (STZ). Normal rat kidney tubular epithelial cells (NRK-52E) were treated with high glucose to induce DN in vitro. Real-time quantitative polymerase chain reaction (RT-qPCR), western blot, immunofluorescence assays were conducted to measure the expression of NUP160, autophagy-associated proteins, and inflammatory cytokines in vitro and in vivo. Pathological changes of kidney and liver tissues were analyzed using hematoxylin and eosin (H&E), Masson and periodic acid-silver (PAS) staining. The body weight, blood glucose, renal and lipid profiles of DN mice were examined. In this study, DN mice showed serious pathological injury. NUP160 expression was upregulated, autophagy was inhibited, and inflammatory response was increased in DN mice. Depletion of NUP160 restored autophagy and inhibited inflammation and fibrosis in high glucose (HG)-treated NRK-52E cells and STZ-induced DN mice by downregulating the expression of p62 and Collagen IV (Col-Ⅳ), increasing the ratio of LC3II/LC3I, and inactivating nuclear factor (NF)-κB signaling. Moreover, NUP160 knockdown could ameliorate pathological damage and glucose tolerance in DN mice. Overall, this study is the first to demonstrate the key role of NUP160 silencing in promoting autophagy against diabetic injury in DN.

Analysis of the Relationships between Mean Red Blood Cell Volume, Red Blood Cell Distribution Width and Hypotension in Patients with Hemodialysis.

BACKGROUND: We aimed to explore and analyze the relationships between mean corpuscular volume (MCV), red blood cell distribution width (RDW) and hypotension in patients with hemodialysis. METHODS: The clinical data of 163 patients from the Xinghua People's Hospital, Taizhou, China with hemodialysis were retrospectively analyzed. The incidence of hypotension was counted and the levels of MCV and RDW were compared between the patients with and without hemodialysis. MCV and RDW were analyzed as possible influencing factors of hypotension. Receiver operating characteristic curve (ROC) was drawn to analyze the effect of MCV and RDW on the risk assessment of hypotension in patients with hemodialysis. RESULTS: MCV in patients with hypotension was significantly lower than those without hypotension (P < 0.05), and RDW was higher than those without hypotension (P < 0.05). The constituent ratio of higher age (>60), diabetic nephropathy, maintenance hemodialysis, MCV < 80fl, RDW > 14.8%, malnutrition, anemia, ultra-filtration rate, diet during dialysis, coronary heart disease, atrial fibrillation and antihypertensive drugs before dialysis were higher in patients with hypotension than those without hypotension (P < 0.05). The sensitivity, specificity and AUC of the combination of MCV and RDW were higher than those of the single assessment. MCV is lower in patients with hypotension and RDW is higher than those in patients without hypotension. CONCLUSION: MCV combined with RDW has a good evaluation effect.

Inhibition of miR-24 suppresses malignancy of human non-small cell lung cancer cells by targeting WWOX in vitro and in vivo.

BACKGROUND: We investigated the effect of micro-RNA 24 (miR-24) and WWOX on non-small cell lung cancer (NSCLC) cell proliferation and migration in vitro and in vivo. METHODS: We performed bioinformatics analysis and 3' untranslated region luciferase assay to investigate the direct target of miR-24. Proliferation, apoptosis, and transwell invasion assays were employed to evaluate the effect of WWOX overexpression with pcDNA3-WWOX and knocking down miR-24 with miR-24 small interfering RNA. Quantitative real-time PCR, Western blot, and immunohistochemistry were also used to investigate miR-24 and c-Kit expression, and apoptosis and invasion-related proteins. Finally, we constructed a tumor xenograft model in nude mice to confirm the effect of miR-24 on NSCLC cell proliferation in vivo. RESULTS: According to our experimental data, miR-24 inhibition could induce apoptosis by activating caspase 3 and suppress the viability and proliferation of NSCLC cells in vitro and in vivo. MiR-24 downregulation could reduce the invasive ability of NSCLC cells by downregulating MMP9. WWOX was identified as a functional target of miR-24. WWOX overexpression generated the same effect with antagonizing miR-24, while blocking WWOX counteracted the tumor suppressive effect caused by miR-24 inhibition. MiR-24 may function as an oncogene and play an important role in the cell growth and migration of NSCLC. CONCLUSIONS: Our findings enhance understanding of the miR-24 regulatory network and the molecular mechanism that underlies the oncogenesis and development of NSCLC. Suppressing the effect of miR-24 on cancer cells using a miR-24 inhibitor may be an attractive therapeutic strategy against NSCLC.

Integrated TCGA analysis implicates lncRNA CTB-193M12.5 as a prognostic factor in lung adenocarcinoma.

BACKGROUND: Lung cancer is a malignant tumor with the highest incidence and mortality around the world. Recent advances in RNA sequencing technology have enabled insights into long non-coding RNAs (lncRNAs), a previously largely overlooked species in dissecting lung cancer pathology. METHODS: In this study, we used a comprehensive bioinformatics analysis strategy to identify lncRNAs closely associated with lung adenocarcinoma, using the RNA sequencing datasets collected from more than 500 lung adenocarcinoma patients and deposited at The Cancer Genome Atlas (TCGA) database. RESULTS: Differential expression analysis highlighted lncRNAs CTD-2510F5.4 and CTB-193M12.5, both of which were significantly upregulated in cancerous specimens. Moreover, network analyses showed highly correlated expression levels of both lncRNAs with those of differentially expressed protein-coding genes, and suggested central regulatory roles of both lncRNAs in the gene co-expression network. Importantly, expression of CTB-193M12.5 showed strong negative correlation with patient survival. CONCLUSIONS: Our study mined existing TCGA datasets for novel factors associated with lung adenocarcinoma, and identified a largely unknown lncRNA as a potential prognostic factor. Further investigation is warranted to characterize the roles and significance of CTB-193M12.5 in lung adenocarcinoma biology.

Correlation Analysis between Obstructive Sleep Apnea Syndrome (OSAS) and Heart Rate Variability.

BACKGROUND: Heart rate variability (HRV) represents the sympathetic nervous system activity induced by apnea or hypopnea events among OSAS patients. However, few studies have been conducted to clarify the association between HRV parameters and polysomnography (PSG) diagnostic indices. In our study, we postulate that the prevalence of cardiac arrhythmias is associated with OSAS, and HRV parameters may be an effective method for OSAS screening. METHODS: Overall, 168 participants had been collected from 2011 to 2016 in the Second Affiliated Hospital of Soochow University. By apnea-hypopnea index (AHI), patients were separated into three subsets: AHI < 5 as control group, 5≤AHI<30 as mild-moderate OSAS group and AHI≥30as severe OSAS group. HRV and PSG parameters were collected based on electrocardiography and polysomnography system. Correlation analyses between standard deviation of R-R intervals (SDNN), SDNN index, RMSSD, PNN50, low frequency (LF), high frequency (HF) and LF/HF ratio and the AHI, ODI and MI were performed by Spearman's correlation analysis. RESULTS: Compared with control group (64.5%) or mild-moderate OSAS group (67.3%), the prevalence of arrhythmias was considerably greater in severe OSAS group (P<0.05). Moreover, we demonstrated that LF/HF was greater in two OSAS groups than the normal group. CONCLUSION: Correlation analyses revealed a significant and positive relation between the LF/HF and AHI, ODI and MI in OSAS patients. Severe OSAS could be attributed to enhanced danger of incident arrhythmia. LF/HF ratio as a relevant feature may be an effective parameter for detecting OSAS.

Expression of miR-106b-25 induced by salvianolic acid B inhibits epithelial-to-mesenchymal transition in HK-2 cells.

Epithelial-to-mesenchymal transition (EMT) is a highly conserved physiological program involved in renal fibrosis. Previous studies have shown that transforming growth factor (TGF)-ß1 induces EMT in human kidney proximal tubular epithelial cells (HK-2 cells), whereas salvianolic acid B (Sal B) has a protective effect against EMT. The molecular pathogenesis of such processes is currently not well understood. In this study, a miRCURYTM LNA Array was used to screen HK-2 cells for expression changes of microRNAs (miRNAs) implicated in EMT. After validation by real-time PCR, all three members of the miR-106b-25 cluster (miR-106b, miR-93, and miR-25) were found to be markedly down-regulated during EMT in response to TGF-ß1, whereas these miRNAs were up-regulated by Sal B treatment in a dose-dependent manner. Moreover, enhanced expression of miR-106b attenuated EMT by retaining the epithelial morphology of HK-2 cells, reducing the levels of α-smooth muscle actin (α-SMA), and increasing the levels of E-cadherin. To explore the molecular basis underlying the inhibitive effect of the miR-106b-25 cluster against EMT, bioinformatics analysis revealed that TGF-ß type II receptor, a regulator of TGF-ß signaling, might be a direct target of the miR-106b-25 cluster. In turn, low levels of TGF-ß type II receptor in EMT of HK-2 cells were shown under the increase of miR-106b. In conclusion, our data suggest that the miR-106b-25 cluster may contribute to EMT in the kidney, and is involved in the protective effect of Sal B. Targeting of specific miRNAs may be a novel therapeutic approach to treat renal fibrosis.