Overexpression of miR-96 leads to retinal degeneration in mice.

Double knockout of miR-183 and miR-96 results in retinal degeneration in mice; however, single knockout of miR-96 leads to developmental delay but not substantial retinal degeneration. To further explore the role of miR-96, we overexpressed this miRNA in mouse retinas. Interestingly, we found that overexpression of miR-96 at a safe dose results in retinal degeneration in the mouse retina. The retinal photoreceptors dramatically degenerated in the miR-96-overexpressing group, as shown by OCT, ERG and cryosectioning at one month after subretinal injection. Degenerative features such as TUNEL signals and reactive gliosis were observed in the miR-96-overexpressing retina. RNA-seq data revealed that immune responses and microglial activation occurred in the degenerating retina. Further qRT‒PCR and immunostaining experiments verified the microglial activation. Moreover, the number of microglia in the miR-96-overexpressing retinas was significantly increased. Our findings demonstrate that appropriate miR-96 expression is required for mouse retinal homeostasis.

Piezo1-dependent regulation of pericyte proliferation by blood flow during brain vascular development.

Blood flow is known to regulate cerebrovascular development through acting on vascular endothelial cells (ECs). As an indispensable component of the neurovascular unit, brain pericytes physically couple with ECs and play vital roles in blood-brain barrier integrity maintenance and neurovascular coupling. However, it remains unclear whether blood flow affects brain pericyte development. Using in vivo time-lapse imaging of larval zebrafish, we monitored the developmental dynamics of brain pericytes and found that they proliferate to expand their population and increase their coverage to brain vessels. In combination with pharmacological and genetic approaches, we demonstrated that blood flow enhances brain pericyte proliferation through Piezo1 expressed in ECs. Moreover, we identified that EC-intrinsic Notch signaling is downstream of Piezo1 to promote the activation of Notch signaling in pericytes. Thus, our findings reveal a role of blood flow in pericyte proliferation, extending the functional spectrum of hemodynamics on cerebrovascular development.

Analysis of Nucleoporin 107 Overexpression and Its Association with Prognosis and Immune Infiltration in Lung Adenocarcinoma by Bioinformatics Methods.

Background: Lung adenocarcinoma (LUAD) has high morbidity and mortality. Current studies indicate nucleoporin 107 (NUP107) is involved in the construction of nuclear pore complex, and NUP107 overexpression contributes to the growth and development in most types of cancers, but its effect in LUAD has not been elucidated. Methods: Differences in NUP107 expression were investigated using the Cancer Genome Atlas (TCGA) and multiple Gene Expression Omnibus (GEO) data sets. Enrichment analysis were implemented to probe the NUP107 function. The association of NUP107 with the degree of immune cell infiltration was investigated by the TIMER database, single-sample gene set enrichment analysis (ssGSEA), and ESTIMATE. The association of NUP107 expression with tumor mutation burden (TMB), TP53, and immune checkpoint was analyzed. Single-cell RNA sequencing data were used to detect NUP107 expression in different cell clusters. Finally, we performed real-time quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry (IHC) to prove the difference of NUP107 expression. Results: NUP107 was overexpressed in LUAD and mainly expressed in cancer stem cell (CSC). Overexpression of NUP107 in LUAD suggested a poorer prognosis. Functional enrichment analysis pointed out that NUP107 was mainly linked to the regulation of cell cycle. Both immune cell infiltration and TMB were found to be in connection with NUP107. Cases in the group with high NUP107 expression had poorer immune infiltration, but had higher expression of immune checkpoints, TMB, and proportion of TP53 mutations. Conclusion: NUP107 is a sensitive diagnostic and prognostic factor for LUAD and may be involved in tumor progression through its effects on cell cycle and immune infiltration.

Photodegradation of amoxicillin by catalyzed Fe3+/H2O2 process.

Three oxidation processes of UV-Fe3+(EDTA)/H2O2 (UV: ultraviolet light; EDTA: ethylenediaminetetraacetic acid), UV-Fe3+/H2O2 and Fe3+/H2O2 were simultaneously investigated for the degradation of amoxicillin at pH 7.0. The results indicated that, 100% amoxicillin degradation and 81.9% chemical oxygen demand (COD(Cr)) removal could be achieved in the UV-Fe3+ (EDTA)/H2O2 process. The treatment efficiency of amoxicillin and COD(Cr) removal were found to decrease to 59.0% and 43.0% in the UV-Fe3+/H2O2 process; 39.6% and 31.3% in the Fe3+/H2O2 process. Moreover, the results of biodegradability (biological oxygen demand (BOD5)/COD(Cr) ratio) revealed that the UV-Fe3+ (EDTA)/H2O2 process was a promising strategy to degrade amoxicillin as the biodegradability of the effluent was improved to 0.45, compared with the cases of UV-Fe3+/H2O2 (0.25) and Fe3+/H2O2 (0.10) processes. Therefore, it could be deduced that EDTA and UV light performed synergetic catalytic effect on the Fe3+/H2O2 process, enhancing the treatment efficiency. The degradation mechanisms were also investigated via UV-Vis spectra, and high performance liquid chromatography-mass spectra. The degradation pathway of amoxicillin was further proposed.

Silencing the expression of lncRNA SNHG15 may be a novel therapeutic approach in human breast cancer through regulating miR-345-5p.

Background: Long noncoding RNA (lncRNA) short nucleolar RNA host gene 15 (SNHG15) has been found to have an oncogenic function in numerous malignancies. Nevertheless, the biological function and regulatory mechanisms of SNHG15 in breast cancer have not been fully elucidated. Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of SNHG15 and in MDA-MB-231 breast cancer cells. The expression of SNHG15 was silenced using small interfering RNA (siRNA) technology. The proliferation and migration of the cells were examined by colony formation assays, cell counting kit 8 (CCK-8) assays, and transwell assays. For the zebrafish xenograft injection experiments, cultured cells labelled with the fluorescent dye CM-DiI were injected into the perivitelline space of the larvae. Results: This present study revealed that the expression of lncRNA SNHG15 (lnc-SNHG15) was significantly upregulated in breast cancer cells, and its overexpression was associated with the tumor. The relative expression of lnc-SNHG15 could be downregulated using siRNAs, and silencing lnc-SNHG15 inhibited the proliferation and the migration of MDA-MB-231 cells. In vivo experiments using the zebrafish xenograft model showed similar results. Mechanistically, the knockdown effect of lnc-SNHG15 could be restored by inhibiting the expression of the miR-345-5p, confirming the negative regulation between lnc-SNHG15 and miR-345-5p. Interestingly, cisplatin treatment combined with SNHG15 knockdown effectively inhibited MDA-MB-231 cell proliferation and migration in the zebrafish xenograft compared to negative controls. Conclusions: In conclusion, lnc-SNHG15 knockdown increased miR-345-5p expression and negated cisplatin resistance in breast cancer cells, and thus, lnc-SNHG15 may be a potential novel target for breast cancer therapy.

Effects of earthworms on surface clogging characteristics of intermittent sand filters.

Intermittent sand filters (ISFs) are effective and economical in treating wastewater, but they are easy to clog up. To explore a feasible and simple method to alleviate clogging, two pilot-scale ISFs were constructed, one of which contained earthworms and the other did not. During the operation, the effects of earthworms on the hydraulic behaviour of ISFs were investigated. The results showed that both ISFs exhibited good performance in wastewater treatment. However, they showed different hydraulic characteristics although operated at the same organic loading rate (approximately 300 g m(-2) d(-1)). The ISF without earthworms clogged only after 53 d operation, and was partially recovered after 7 d resting, but after that, clogging occurred again, and more rapidly than the initial clogging event (40 d). However, water on the medium surface of the ISF with earthworms was not observed during the whole experiments. In addition, 11-13% of effective porosity and 0.015-0.026 cm s(-1) of infiltration rate were measured in the upper 20 cm of the ISF at the end of the experiments. The facts demonstrated that earthworms played a positive role in alleviating clogging and earthworms fed filter could alleviate surface clogging effectively.

Excess nitrogen accumulation in activated sludge in sequencing batch reactor with a single-stage oxic process.

It was occasionally found that a significant nitrogen loss in solution under neutral pH value in a sequencing batch reactor with a single-stage oxic process using synthetic wastewater, and then further studies were to verify the phenomenon of nitrogen loss and to investigate the pathway of nitrogen removal. The result showed that good performance of nitrogen removal was obtained in system. 0-7.28 mg L(-1) ammonia, 0.08-0.38 mg L(-1) nitrite and 0.94-2.12 mg L(-1) nitrate were determined in effluent, respectively, when 29.85-35.65 mg L(-1) ammonia was feeding as the sole nitrogen source in influent. Furthermore, a substantial nitrogen loss in solution (95% of nitrogen influent) coupled with a little gaseous nitrogen increase in off-gas (7% of nitrogen influent) was determined during a typical aerobic phase. In addition, about 322 mg nitrogen accumulation (84% of nitrogen influent) was detected in activated sludge. Based on nitrogen mass balance calculation, the unaccounted nitrogen fraction and the ratio of nitrogen accumulation in sludge/nitrogen loss in solution were 14.6 mg (3.7% of nitrogen influent) and 0.89, respectively. The facts indicated that the essential pathway of nitrogen loss in solution in this study was excess nitrogen accumulation in activated sludge.

EDTA and electricity synergetic catalyzed Fe(3+)/H2O2 process for amoxicillin oxidation.

Three oxidation processes for amoxicillin wastewater pretreatment such as Electro-Fe(3+)(EDTA)/H2O2 (EDTA: ethylenediaminetetraacetic acid), Fe(3+)(EDTA)/H2O2 and Electro-Fe(3+)/H2O2 were simultaneously discussed at pH of 7.0 (+/-0.1). It was found that the above processes could achieve 78%, 64%, 33% chemical oxygen demand (COD(cr)) removal, and 86%, 70%, 47% amoxicillin degradation respectively. Moreover, the results of biodegradability (biological oxygen demand (BOD(5))/COD(cr) ratio) showed that the Electro-Fe(3+)(EDTA)/H2O2 process was a promising way to pretreat antibiotic wastewater due to the biodegradability of the effluent improved to 0.48 compared with the cases of Fe(3+)(EDTA)/H2O2 (0.40) and Electro-Fe(3+)/H2O2 process (0.12). Therefore, it was reasonable to note that EDTA and electricity showed synergetic effect on the oxidation process. Additionally, infrared spectra (IR) were applied to concisely propose a potential degradation way of amoxicillin. The characteristic changes of H2O2 and EDTA in the oxidation process were also investigated in detail.

Protective effect of dexmedetomidine against diabetic hyperglycemia-exacerbated cerebral ischemia/reperfusion injury: An in vivo and in vitro study.

AIMS: Dexmedetomidine (Dex) has been noted to have neuroprotective effect against cerebral ischemia-reperfusion (I/R) injury. However, the effect of Dex in diabetic hyperglycemia-exacerbated cerebral I/R injury and its underlying mechanism remain unclear. MAIN METHODS: The infarct volume and brain edema were evaluated by 2,3,5-triphenyltetrazolium chloride staining and standard wet-dry method. Modified neurological severity score was utilized to assess the neurological deficits. The oxidative stress and inflammation were evaluated by detecting reactive oxygen species (ROS), malondialdehyde (MDA), tumor necrosis factor (TNF)-α and interleukin (IL)-1ß. Terminal deoxynucleotidyl transferase dUTP nick end labeling assay and cell count kit-8 were applied to measure cell apoptosis and viability. KEY FINDINGS: Dex treatment reduced infarct volume, decreased brain water content and improved neurological deficit in middle cerebral artery occlusion/reperfusion (MCAO/R) mice. Dex treatment reduced the levels of ROS, MDA, TNF-α and IL-1ß in the entire middle cerebral artery territory of diabetic mice subjected to MCAO/R, as well as in primary culture of mouse hippocampal neurons stimulated with 50 mM glucose and oxygen glucose deprivation/reperfusion. Dex treatment inhibited neuronal apoptosis induced by diabetic hyperglycemia-exacerbated cerebral I/R injury. Dex upregulated nuclear factor of activated T-cells 5 (NFAT5) and Sirtuin 1 (SIRT1) expression, induced NF-E2-related factor 2 (Nrf2) translocation from cytoplasm to nucleus and inhibited the acetylation of Nrf2. However, these changes triggered by Dex treatment were abrogated by NFAT5 knockdown. SIGNIFICANCE: Dex protects against diabetic hyperglycemia-exacerbated cerebral I/R injury through attenuation of oxidative stress, inflammation and apoptosis. The underlying mechanism is at least the NFAT5/SIRT1/Nrf2 signaling pathway dependent.

MicroRNA-137 and its downstream target LSD1 inversely regulate anesthetics-induced neurotoxicity in dorsal root ganglion neurons.

PURPOSE: Anesthetic reagents, such as bupivacaine (Bv), induce significant neurotoxicity in dorsal root ganglion neurons (DRGNs). In this study, we investigated the expression, function and cross-association of microRNA-137-3p (miR-137-3p) and lysine (K)-specific demethylase 1A (LSD1) in a murine model of Bv-induced neural injury in DRGNs. METHODS: Murine DRGNs were culture in vitro and treated with Bv. QPCR was used to evaluate miR-137-3p expression in Bv-injured DRGNs. MiR-137-3p was genetically downregulated to evaluate its rescuing effect on Bv-induced DRGN apoptosis and neurite retraction. The association of miR-137-3p on its downstream target, LSD1 coding gene KDM1A, was evaluated by dual-luciferase activity assay and qPCR. In miR-137-3p-downregulated DRGNs, KDM1A was inhibited to evaluate its involvement in miR-137-3p-mediated modulation on Bv-induced DRGN neurotoxicity. Furthermore, KDM1A expression in Bv-injured DRGN was evaluated by qPCR, and LSD1 was overexpressed in DRGN to evaluate its direct effect on Bv-induced neurotoxicity. RESULTS: MiR-137-3p was upregulated in Bv-injured DRGNs. MiR-137-3p downregulation rescued Bv-induced DRGN apoptosis and neurite retraction. LSD1 was demonstrated to be downstream to, and inversely modulated by miR-137-3p in DRGN. In Bv-injured DRGNs, LSD1 downregulation reversed miR-137-3p-downregualtion-induced neural protection. Furthermore, LSD1 upregulation directly rescued Bv-induced apoptosis and neurite retraction in DRGNs. CONCLUSIONS: MiR-137-3p and its downstream target LSD1 are inversely associated to regulate anesthetics-induced neurotoxicity in DRGN. This signaling pathway may be a therapeutic candidate to reduce anesthetics-induced neurological damage in human patients.

Co-Positivity for Anti-dsDNA, -Nucleosome and -Histone Antibodies in Lupus Nephritis Is Indicative of High Serum Levels and Severe Nephropathy.

OBJECTIVE: To characterize the significance of correlated autoantibodies in systemic lupus erythematosus (SLE) and its complication lupus nephritis (LN) in a large cohort of patients. METHODS: Clinical data were statistically analyzed in 1699 SLE patients with or without nephritis who were diagnosed and treated during 2002-2013 in the northeast region of China. Reactivity to a list of 16 autoantibodies was detected by the serum test Euroline ANA profile (IgG). Serum titers of the anti-nucleosome autoantibodies were measured by ELISA assays. Kidney biopsies were examined by pathologists. Immune complex deposition was identified by immunohistochemistry stain. RESULTS: Simultaneous positivity of anti-dsDNA, -nucleosome and -histone antibodies (3-pos) was prevalent in SLE patients with LN compared to Non-renal SLE patients (41% vs 11%, p< 0.001). Significant correlations were found between any two of the above three anti-nucleosome antibodies in LN patients. In comparison to non-3-pos cohorts, 3-pos patients with LN had significantly higher serum levels of the three antibodies and more active disease; was associated with type IV disease; suffered from more severe renal damages; received more intensive treatment and had worse disease outcome. The serum levels of these three autoantibodies in 3-pos LN patients were significantly decreased when they underwent clinical recovery. CONCLUSIONS: Simultaneous reactivity to anti-dsDNA, -nucleosome and -histone antibodies by Euroline ANA profile (IgG) may indicate severe nephropathy in patients with SLE.

The probable metabolic relation between phosphate uptake and energy storages formations under single-stage oxic condition.

To investigate the possible biochemical metabolisms for excess phosphate uptake in a sequencing batch reactor (SBR) with single-stage oxic process, which was reported using glucose as the sole carbon source previously, glucose and acetate were fed to two SBRs as the sole carbon source, respectively. The changes of polyhydroxyalkanoates (PHAs), glycogen and the removal of phosphorus were compared between two SBRs. It was observed that the phosphorus removal efficiency was 91.8-94.4% with glucose, and 23.3-28.5% with acetate, although the former showed much lower accumulations/transformations of PHAs. Instead, the former showed a much higher transformation of glycogen. The facts suggested that glycogen could replace PHAs to supply energy for phosphate uptake under the single-stage oxic condition. Furthermore, the possible biochemical metabolisms were proposed to describe the relation between phosphate uptake and energy storages formations under such a single-stage oxic process. Such a process may serve as a prototype for the development of alternative biological and chemical options for phosphate removal from wastewaters.