Circ-Luc7l Absence Attenuates Diabetic Nephropathy Progression by Reducing Mesangial Cell Excessive Proliferation, Inflammation, and Extracellular Matrix Accumulation via Mediating the miR-205-5p/Tgfbr1 Pathway.

Diabetic nephropathy (DN) threatens the survival quality of patients, with complex pathogenesis. Circular RNA (circRNA) dysregulation occurs in DN development. This work aimed to investigate the role of circ-Luc7l in DN cell models and related molecular mechanisms. The expression of circ-Luc7l, microRNA (miR)-205-5p, and transforming growth factor-beta receptor 1 (Tgfbr1) was examined by real-time quantitative PCR (RT-qPCR). Cell viability and proliferation were detected by Cell Counting Kit-8 (CCK-8) assay and EdU assay. The expression of extracellular matrix (ECM)-related markers and Tgrbr1 protein was measured by Western blot. The binding between miR-205-5p and circ-Luc7l or Tgfbr1 was validated by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay, or RNA pull-down assay. Experimental animal models were established to elucidate the function of circ-Luc7l in vivo. Circ-Luc7l expression was notably enhanced in high glucose (HG)-treated mesangial cells. Knockdown of circ-Luc7l attenuated HG-induced cell proliferation, inflammation, and ECM accumulation in vitro and relieved inflammation and ECM accumulation of kidneys of diabetic mice in vivo. Circ-Luc7l targeted miR-205-5p, and miR-205-5p inhibition rescued the depletion effects of circ-Luc7l knockdown on cell proliferation, inflammation, and ECM accumulation. MiR-205-5p bound to Tgfbr1 whose expression was negatively regulated by circ-Luc7l. Tgfbr1 overexpression also rescued the depletion effects of circ-Luc7l knockdown on cell proliferation, inflammation, and ECM accumulation. In HG conditions, increased circ-Luc7l upregulated Tgfbr1 expression via targeting miR-205-5p to induce DN progression.

Disrupting circ-GNB4 mitigates high glucose-induced human mesangial cells injury by regulating the proliferation, ECM accumulation, inflammation and oxidative stress through circ-GNB4/miR-23c/EGR1 pathway [RETRACTED].

ABSTRACT: Diabetic nephropathy (DN) is the most common complication of diabetes mellitus. Although G protein subunit beta 4 (GNB4)-derived circular RNA (circ-GNB4; hsa_circ_0068087) is a promising candidate biomarker in diabetes mellitus, whether circ-GNB4 participates in DN occurrence and development remains unknown. Herein, we focused on DN-associated human renal mesangial cells (HRMCs) injury, and HRMCs were exposed in high glucose (HG) condition. Using quantitative polymerase chain reaction and western blotting, we found that circ-GNB4 and early growth response factor 1 (EGR1) were upregulated, whereas microRNA (miR)-23c was downregulated in DN patients' sera and HG-stimulated HRMCs. HG-induced injuries were measured by MTS method, western blotting, enzyme-linked immunosorbent assay and other special assay kits. Consequently, HG could inhibit superoxide dismutase activity, but induce cell proliferation and levels of malondialdehyde, Fibronectin, Collagen I, Collagen IV, interleukin-6, interleukin-1ß, and tumor necrosis factor-α. However, HG-induced these injuries were overall suppressed by silencing circ-GNB4 or overexpressing miR-23c. Moreover, miR-23c knockdown could counteract the effect of circ-GNB4 deficiency, and EGR1 restoration abrogated miR-23c overexpression role in HG-stimulated HRMCs. Notably, circ-GNB4 could target miR-23c and EGR1 was targeted by miR-23c, as confirmed by dual-luciferase reporter assay and RNA immunoprecipitation. Moreover, EGR1 expression was positively modulated by circ-GNB4 via miR-23c. Collectively, circ-GNB4 might be a novel mechanism of DN-induced HRMCs injury, and there was a circ-GNB4/miR-23c/EGR1 pathway underlying the proliferation, extracellular matrix accumulation, inflammation and oxidative stress. This study suggested circ-GNB4 as a potential target to interfere the development of DN.

Resveratrol Reduces Kidney Injury in a Rat Model of Uremia and is Associated with Increased Expression of Heat Shock Protein 70 (Hsp70).

BACKGROUND This study aimed to investigate the effects of resveratrol on kidney function in a rat model of uremia and the expression of heat shock proteins. MATERIAL AND METHODS The rat model of uremia was developed by 5/6 nephrectomy of Sprague-Dawley rats. The Hsp70 inhibitor MKT-077, a rhodacyanine dye, was used. The study groups included rats with sham surgery (the sham group), the rat model of uremia (the model group), the solvent-treated control group (the control group), the rat model treated with resveratrol group (the resveratrol group), the rat model treated with MKT-077 (the MKT-077 group), and the resveratrol+MKT-077 group. Kidney tissues were studied histologically. Renal cell apoptosis was detected by the TUNEL method. Expression of p53, Bax, and Bcl-2 mRNA and protein were detected by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and immunohistochemistry, respectively. RESULTS Compared with the sham group, the expression levels of heat shock proteins Hsp70, Hsp90, Hsp27, Hsp25, Hsp40, and Hsp60 in the kidney of the rat model group increased to different degrees. Compared with the model group, the Hsp70 levels in the resveratrol group were significantly increased (p<0.05). Compared with the model group, treatment with MKT-077 reduced the survival rate of rats, which was increased following resveratrol treatment. Compared with the resveratrol group, renal function in the resveratrol+MKT-077 group was significantly reduced (p<0.05). CONCLUSIONS In a rat model of uremia, resveratrol reduced renal injury and improved both renal function and survival, which were associated with increased expression of Hsp70.

Saturable absorber based on a single mode fiber - graded index fiber - single mode fiber structure with inner micro-cavity.

An Er-doped mode-locked fiber laser with a saturable absorber based on single mode - graded index multimode - single mode fiber (SMF-GIMF-SMF) with inner micro-cavity is demonstrated. The modulation depth of the saturable absorber was measured to be 1.9% when the SMF-GIMF-SMF structure is bent to a certain state. Such a simple saturable absorber enables the mode-locking operation in a ring Er-doped fiber laser and ultrafast pulses with pulse energy of 0.026 nJ and pulse width of 528 fs at the fundamental repetition rate of 14.34 MHz can be generated. In addition, the harmonic mode-locking operation can also be achieved.

Stretched graded-index multimode optical fiber as a saturable absorber for erbium-doped fiber laser mode locking.

A novel mode-locking method based on the nonlinear multimode interference in the stretched graded-index multimode optical fiber (GIMF) is proposed in this Letter. The simple device geometry, where the light is coupled in and out of the stretched GIMF via single-mode fibers, is demonstrated to exhibit the temporal intensity discrimination required for mode locking. The nonlinear saturable absorber (SA) characteristics of the device are controllable by simply adjusting the strength of the stretching applied. The modulation depth of the device, which consists of ∼23.5 cm GIMF, is tuned from 10.37% to 22.27%. Such a simple SA enables the wavelength-switchable mode-locking operation in a ring Er-doped fiber laser, and ultrafast pulses with a pulse width of 506 fs at 1572.5 nm and 416 fs at 1591.4 nm were generated. The versatility and simplicity of the SA device, together with the possibility of scaling the pulse energy, make it highly attractive in ultrafast photonics.

Effect of Vitamin D and Tacrolimus Combination Therapy on IgA Nephropathy.

BACKGROUND To explore the effects and the mechanism of vitamin D (VD) and tacrolimus (TAC) combinatorial therapy in the treatment of IgA nephropathy (IgAN) in a rat model. MATERIAL AND METHODS IgAN rat models constructed by oral immunization with bovine serum albumin (BSA) and lipopolysaccharide (LPS) (n=30) and were treated with: saline (model group), TAC (TAC group), or TAC+VD therapy (TAC+VD group) through gavage daily for 14 days. Serum creatinine (Scr), albumin (ALB), blood urea nitrogen (BUN), and urinary protein (UAE) levels were determined. Histopathology of renal tissues was examined after hematoxylin and eosin (H&E) staining. The levels of cytokines TGF-ß1, IL-5, IFN-γ, and IL-4 in serum were detected by enzyme-linked immunosorbent assay (ELISA). Changes in TLR4/NF-κB pathway were evaluated by western blot. RESULTS Both TAC and TAC+VD treatment significantly restored the dysregulated Scr, ALB, BUN, and UAE levels in IgAN rats. TAC+VD therapy more prominently restored Scr and UAE levels (p<0.05). TAC+VD therapy demonstrated superior efficacy in reducing glomerular mesangial cells hyperplasia, reducing thickening of the glomerular basement membrane and glomerular infiltration of inflammatory cells. Thymus and spleen indexes were also increased (p<0.05). The levels of TGF-ß1, IL-5, and IL-4 of the TAC+VD group were also lower than those of the TAC group (p<0.05). The TAC+VD group also demonstrated increased IFN-γ, and decreased p-P65/P65 and TLR4 compared to the TAC group. CONCLUSIONS TAC+VD combinatorial therapy can effectively alleviate renal tissue damage in IgAN rats by regulating immune response and the NF-κB/TLR4 pathway.

Hierarchical Ohmic Contact Interface Engineering for Efficient Hydrazine-Assisted Hydrogen Evolution Reaction.

Hydrazine oxidation reaction coupled with hydrogen evolution reaction (HER) is an effective strategy to achieve low energy water splitting for hydrogen production. In order to realize the application of hydrazine-assisted HER system, researchers have been focusing on the development of electrocatalysts with integrated dual active sites, while the performance under high current density is still unsatisfying. In this work, hierarchical Ohmic contact interface engineering is designed and used as a bridge between the NiMo and Ni2 P heterojunction toward industrial current density applications, with the charge transfer impedance greatly eliminated via such a pathway with low energy barrier. As a proof-of-concept, the importance of charge redistribution and energy barrier at the Ohmic contact interface is investigated by significantly reducing the voltage of overall hydrazine splitting (OHzS) at high current density. Intriguingly, the NiMo/Ni2 P hierarchical Ohmic contact heterojunction can drive current densities of 100 and 500 mA cm-2 with only 181 and 343 mV cell voltage in the OHzS electrolyzer with high electrocatalytic stability. The proposed hierarchical Ohmic contact interface engineering paves new avenue for hydrogen production with low energy consumption.

The discovery of an anti-Candida xanthone with selective inhibition of Candida albicans GAPDH.

OBJECTIVES: This study aimed to discover novel antifungals targeting Candida albicans glyceraldehyde-3-phosphate dehydrogenase (CaGAPDH), have an insight into inhibitory mode, and provide evidence supporting CaGAPDH as a target for new antifungals. METHODS: Virtual screening was utilized to discover inhibitors of CaGAPDH. The inhibitory effect on cellular GAPDH was evaluated by determining the levels of ATP, NAD, NADH, etc., as well as examining GAPDH mRNA and protein expression. The role of GAPDH inhibition in C. albicans was supported by drug affinity responsive target stability and overexpression experiments. The mechanism of CaGAPDH inhibition was elucidated by Michaelis-Menten enzyme kinetics and site-specific mutagenesis based on docking. Chemical synthesis was used to produce an improved candidate. Different sources of GAPDH were used to evaluate inhibitory selectivity across species. In vitro and in vivo antifungal tests, along with anti-biofilm activity, were carried out to evaluate antifungal potential of GAPDH inhibitors. RESULTS: A natural xanthone was identified as the first competitive inhibitor of CaGAPDH. It demonstrated in vitro anti-C. albicans potential but also caused hemolysis. XP-W, a synthetic side-chain-optimized xanthone, demonstrated a better safety profile, exhibiting a 50-fold selectivity for CaGAPDH over human GAPDH. XP-W also exhibited potent anti-biofilm activity and displayed broad-spectrum anti-Candida activities in vitro and in vivo, including multi-azole-resistant C. albicans. CONCLUSIONS: These results demonstrate for the first time that CaGAPDH is a valuable target for antifungal drug discovery, and XP-W provides a promising lead.

Scalable Customization of Crystallographic Plane Controllable Lithium Metal Anodes for Ultralong-Lasting Lithium Metal Batteries.

A formidable challenge to achieve the practical applications of rechargeable lithium (Li) metal batteries (RLMBs) is to suppress the uncontrollable growth of Li dendrites. One of the most effective solutions is to fabricate Li metal anodes with specific crystal plane, but still lack of a simple and high-efficient approach. Herein, a facile and controllable way for the scalable customization of polished Li metal anodes with highly preferred (110) and (200) crystallographic orientation (donating as polished Li(110) and polished Li(200), respectively) by regulating the times of accumulative roll bonding, is reported. According to the inherent characteristics of polished Li(110)/Li(200), the influence of Li atomic structure on the electrochemical performance of RLMBs is deeply elucidated by combining theoretical calculations with relative experimental proofs. In particular, a polished Li(110) crystal plane is demonstrated to induce Li+ uniform deposition, promoting the formation of flat and dense Li deposits. Impressively, the polished Li(110)||LiFePO4 full cells exhibit unprecedented cycling stability with 10 000 cycles at 10 C almost without capacity degradation, indicating the great potential application prospect of such textured Li metal. More valuably, this work provides an important reference for low-cost, continued, and large-scale production of Li metal anodes with highly preferred crystal orientation through roll-to-roll manufacturability.

Designing a Stable Solid Electrolyte Interphase on Lithium Metal Anodes by Tailoring a Mg Atom Center and the Inner Helmholtz Plane for Lithium-Sulfur Batteries.

Arising from the extraordinary theoretical energy density, rechargeable lithium-sulfur (Li-S) batteries have been reputed as one of the most appealing options for next-generation high-performance energy storage and conversion devices. Unfortunately, their industrial implementation has been strongly governed by the formation of Li dendrites caused by the unstable solid electrolyte interphase (SEI) film. Herein, we report a novel electrolyte by introducing the Mg(NO3)2 additive to suppress the growth of Li dendrites, further improving the cycling lifetime of Li-S batteries. On the one hand, Mg2+ can rapidly react with Li atoms to generate Mg atoms, replacing the Li atoms on the top surface of Li metal and forming the Mg center simultaneously. On the other hand, NO3- can be adsorbed in the inner Helmholtz plane and reduced as an inorganic-rich SEI film for stabilizing the Li metal anode when the electrolyte comes in contact with Li metal, effectively mitigating the formation of Li dendrites. Combining the experimental results and theoretical calculations, we confirm that the Mg atom center and the inorganic-rich SEI film are both beneficial for enhancing the electrochemical performance of Li-S batteries. This work provides a new insight into the electrolyte additive and a possible alternative for the design of high-performance Li-S batteries beyond the LiNO3 additive.

Anion-Immobilized Gel Polymer Electrolyte with a High Ion Transference Number for High-Performance Lithium/Sodium Metal Batteries.

Due to their high energy density, lithium/sodium metal batteries (LMBs/SMBs) are expected to be the next generation of energy storage systems. However, the further application of alkali metal batteries based on liquid electrolytes is limited due to increasing safety concerns. Gel polymer electrolytes (GPEs), which combine the advantages of the high ionic conductivity of liquid electrolytes and excellent mechanical properties of solid polymer electrolytes, are considered to play an irreplaceable role in the realization of high-performance alkali metal batteries. In this work, a flexible boron-containing GPE (B-GPE) with a cross-linked polymer network structure is prepared by a UV-induced process. The as-prepared B-GPE exhibits good ionic conductivity and has an extremely high ion transference number due to the electron-withdrawing effect of the boron moiety and the facile electrolyte uptake ability of the ethylene oxide chain. Furthermore, a "gentle" electrode/electrolyte contact is designed by a one-step in situ polymerization method, which can enhance ion transport within the electrode and at the electrode/electrolyte interface due to the presence of a continuous polymer phase for ion conduction. Therefore, LMBs and SMBs containing B-GPE are able to effectively inhibit the growth of dendrites while exhibiting excellent cycling stability. These comprehensive results indicate that this novel B-GPE possesses potential applications for high-performance alkali metal batteries.

[ABA signaling mediated by PYR/PYL/RCAR in plants].

Abscisic acid (ABA) is a phytohormone that plays critical roles in numerous developmental stages as well as in adaptive responses to biotic and abiotic stresses. Recent breakthroughs in the field of ABA signaling have indicated that there are three major components, PYR/PYL/RCAR (an ABA receptor), type 2C protein phosphates (PP2C, a negative regulator), and SNF1-related protein kinase 2 (SnRK2, a positive regulator). Further results show that these three proteins construct a double negative regulatory system, PYR/PYL/RCAR-| PP2C-| SnRK2, to regulate ABA signal responses in plant cells. Moreover, the combination patterns of these components in vivo are restricted by spatio-temporal and biochemical determinants and the combinational variation in the ABA signalosome is specific to different ABA signal responses. This review summarizes recent advances of study on the molecular basis and regulatory mechanism of PYR/PYL/RCAR-mediated ABA signaling pathway and PYR/PYL/RCAR-PP2C-SnRK2 complex-mediated ABA signal regulation network in plants. The perspectives related to this study are proposed.

Corrigendum: CircAFF1 aggravates vascular endothelial cell dysfunction mediated by miR-516b/SAV1/YAP1 axis.

[This corrects the article DOI: 10.3389/fphys.2020.00899.].

Role of CD2-associated protein in albumin overload-induced apoptosis in podocytes.

Proteinuria is a well-established exacerbating factor of chronic kidney diseases. However, the harmful effects of protein overload on podocytes and the underlying mechanisms are still poorly understood. In the present study, we examined the effects of high concentrations of albumin on podocytes and investigated the role of CD2AP (CD2-associated protein) in albumin overload-induced podocyte apoptosis. Conditionally immortalized mouse podocytes were cultured in vitro and treated with different concentrations of BSA. In addition, CD2AP eukaryotic expression vector or siRNA (small interfering RNA) was transfected into podocytes before they were exposed to BSA. Podocyte apoptosis, expressions of active caspase-3 (p17) and CD2AP, and the distribution of F-actin cytoskeleton were detected by flow cytometry, Western-blot analysis and fluorescent staining respectively. It was found that exposure of podocytes to BSA induced podocyte apoptosis in a concentration-dependent manner that was accompanied by up-regulation of active caspase-3, the disruption of F-actin cytoskeleton, and decreased expression of CD2AP. Transfection of CD2AP eukaryotic expression vector into podocytes increased CD2AP expression, partially restored F-actin distribution, blocked active caspase-3 expression and inhibited podocyte apoptosis. In contrast, transfection of CD2AP siRNA deteriorated the above changes induced by BSA. It is concluded that protein overload induces podocyte apoptosis via the down-regulation of CD2AP and subsequent disruption of cytoskeleton of podocytes, and CD2AP may play an important role in protein overload-induced podocyte injury.

Protective effects of eplerenone on podocyte injury in adriamycin nephropathy rats.

To investigate the protective effects of eplerenone on adriamycin-induced renal injury and the possible mechanisms involved, 36 male Sprague-Dawley rats were randomly divided into control group, adriamycin nephropathy (AN) group and eplerenone-treated group (100 mg·kg(-1)·d(-1) eplerenone). Blood pressure, 24-h urinary protein, serum potassium, sodium and creatinine were measured 28 days after adriamycin injection (a single tail intravenous injection of 6.5 mg/kg adriamycin). The morphological changes of renal tissues were observed by light and electron microscopy. Immunohistochemistry and Western blotting were performed to examine the expression of TGF-ß(1) and desmin in renal cortex. The results showed that 28 days after adriamycin injection, there were no significant changes in the level of serum potassium, sodium, creatinine concentrations and blood pressure values in the rats of the three groups. Meanwhile, the 24-h proteinuria excretion in the AN group was significantly higher than that in the control group (P<0.01), but that in the eplerenone-treated group was substantially reduced when compared with that in the AN group (P<0.05). Mild mesangial cell proliferation and matrix expansion, diffuse deformation and confluence of foot processes in podocytes were found in the AN group. By contrast, rats in the eplerenone-treated group exhibited obvious attenuation of these morphological lesions. The protein expression of TGF-ß(1) and desmin in the AN group was markedly up-regulated in contrast to that in the control group (P<0.01), whereas that in the eplerenone-treated group was much lower than in the AN group (P<0.05). It was concluded that eplerenone may ameliorate the proteinuria and the development of pathological alteration in adriamycin-induced nephropathy presumably via the inhibition of cytokine release, and restore the morphology of podocytes independent of its blood pressure-lowing effects.

Exhausting T Cells During HIV Infection May Improve the Prognosis of Patients with COVID-19.

T-cell reduction is an important characteristic of coronavirus disease 2019 (COVID-19), and its immunopathology is a subject of debate. It may be due to the direct effect of the virus on T-cell exhaustion or indirectly due to T cells redistributing to the lungs. HIV/AIDS naturally served as a T-cell exhaustion disease model for recognizing how the immune system works in the course of COVID-19. In this study, we collected the clinical charts, T-lymphocyte analysis, and chest CT of HIV patients with laboratory-confirmed COVID-19 infection who were admitted to Jin Yin-tan Hospital (Wuhan, China). The median age of the 21 patients was 47 years [interquartile range (IQR) = 40-50 years] and the median CD4 T-cell count was 183 cells/µl (IQR = 96-289 cells/µl). Eleven HIV patients were in the non-AIDS stage and 10 were in the AIDS stage. Nine patients received antiretroviral treatment (ART) and 12 patients did not receive any treatment. Compared to the reported mortality rate (nearly 4%-10%) and severity rate (up to 20%-40%) among COVID-19 patients in hospital, a benign duration with 0% severity and mortality rates was shown by 21 HIV/AIDS patients. The severity rates of COVID-19 were comparable between non-AIDS (median CD4 = 287 cells/µl) and AIDS (median CD4 = 97 cells/µl) patients, despite some of the AIDS patients having baseline lung injury stimulated by HIV: 7 patients (33%) were mild (five in the non-AIDS group and two in the AIDS group) and 14 patients (67%) were moderate (six in the non-AIDS group and eight in the AIDS group). More importantly, we found that a reduction in T-cell number positively correlates with the serum levels of interleukin 6 (IL-6) and C-reactive protein (CRP), which is contrary to the reported findings on the immune response of COVID-19 patients (lower CD4 T-cell counts with higher levels of IL-6 and CRP). In HIV/AIDS, a compromised immune system with lower CD4 T-cell counts might waive the clinical symptoms and inflammatory responses, which suggests lymphocyte redistribution as an immunopathology leading to lymphopenia in COVID-19.

Resveratrol Plays Protective Roles on Kidney of Uremic Rats via Activating HSP70 Expression.

OBJECTIVE: To investigate the protective effects of resveratrol on kidney of uremic rats and to explore whether the mechanism is associated with heat shock protein 70 (HSP70) expression. METHODS: Sixty male Sprague Dawley rats were randomly separated into 5 groups, including sham group, uremic model group, and different doses of resveratrol group (5 mg/kg, 10 mg/kg, and 20 mg/kg). The serum creatinine (Cr) and urea nitrogen (BUN) levels were detected by Automatic Biochemical Analyzer (ABA). The pathological changes of renal tissues and the renal interstitial fibrosis were analyzed by hematoxylin-eosin (HE) and Masson, respectively. The expression of HSP70 protein in renal tissues was detected by immunohistochemistry. The expression of HSP70 and NF-κB pathway-related proteins were detected by Western blot. To further validate the protective role of resveratrol through activating HSP70 in uremic rats, HSP70 activator (17-AAG) and HSP70 inhibitor group (MKT-077) were used. RESULTS: In the model group, the levels of Cr and BUN in serum were significantly increased, and the renal interstitial collagen deposition was also obviously increased (p < 0.05). Compared with the model group, the levels of Cr and BUN in different doses of resveratrol groups were remarkably declined, and the renal interstitial collagen deposition was declined (p < 0.05). Resveratrol also significantly improved the renal tissue lesions when compared with the model group. In renal tissues, different doses of resveratrol treatment remarkably raised HSP70 and p-IκBα expression and also remarkably declined the level of p-P65 protein (p < 0.05). Meanwhile, the effect of 17-AAG was similar to 20 mg/kg resveratrol on NF-κB pathway-related proteins expression. After the added MKT-077 in the resveratrol treatment group, the levels of HSP70 and p-IκBα in the renal tissue were remarkably declined; however, the levels of p-P65 protein was remarkably raised (p < 0.05). CONCLUSION: Resveratrol played a protective role on the kidney of uremic rats through activating HSP70 expression.

Long non-coding RNA CDKN2B-AS1 regulates high glucose-induced human mesangial cell injury via regulating the miR-15b-5p/WNT2B axis.

BACKGROUND: Long non-coding RNA cyclin-dependent kinase inhibitor 2B antisense RNA 1 (CDKN2B-AS1) has been reported to be related to diabetic nephropathy (DN) progression. However, the regulatory mechanisms of CDKN2B-AS1 in DN are unclear. METHODS: High glucose (HG) was used to induce human mesangial cells (HMCs) for establishing the DN model. Expression levels of CDKN2B-AS1, microRNA (miR)-15b-5p, wingless-Type family member 2B (WNT2B) mRNA in serum and HMCs were detected through quantitative real-time polymerase chain reaction (qRT-PCR). The viability and cell cycle progression of HMCs were determined with Cell Counting Kit-8 (CCK-8) or flow cytometry assays. The levels of several proteins and inflammatory factors in HMCs were analyzed by western blotting or enzyme-linked immunosorbent assay (ELISA). The relationship between CDKN2B-AS1 or WNT2B and miR-15b-5p was verified with dual-luciferase reporter assay. RESULTS: CDKN2B-AS1 and WNT2B were upregulated while miR-15b-5p was downregulated in serum of DN patients and HG-treated HMCs. CDKN2B-AS1 inhibition reduced HG-induced viability, cell cycle progression, ECM accumulation, and inflammation response in HMCs. CDKN2B-AS1 regulated WNT2B expression via competitively binding to miR-15b-5p. MiR-15b-5p inhibitor reversed CDKN2B-AS1 knockdown-mediated influence on viability, cell cycle progression, ECM accumulation, and inflammation response of HG-treated HMCs. The repressive effect of miR-15b-5p mimic on viability, cell cycle progression, ECM accumulation, and inflammation response of HG-treated HMCs was abolished by WNT2B overexpression. CONCLUSION: CDKN2B-AS1 regulated HG-induced HMC viability, cell cycle progression, ECM accumulation, and inflammation response via regulating the miR-15b-5p/WNT2B axis, provided a new mechanism for understanding the development of DN.

Trace analysis of 28 antibiotics in plant tissues (root, stem, leaf and seed) by optimized QuEChERS pretreatment with UHPLC-MS/MS detection.

Phytoremediation has proven to be an effective in-situ treatment technique for antibiotic contamination. Due to the immature methods of extracting multi-antibiotics in different plant tissues, the antibiotic absorption and transportation mechanism in the phytoremediation process has yet to be resolved. Therefore, an improved Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) pretreatment with ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) detection method for 28 antibiotics in different plant tissues (root, stem, leaf and seed) was developed in this study. The optimized method showed satisfactory performance with recoveries for most antibiotics ranging from 70% to 130% (except sulfadoxine with 138 ± 8.84% in root, sulfameter with 68.9 ± 1.87% and sulfadoxine with 141 ± 10.0% in seed). The limits of detection (LODs) of the target compounds in root, stem, leaf and seed were 0.04 ± 0.02 ~ 2.50 ± 1.14 ng/g, 0.05 ± 0.02 ~ 1.78 ± 0.42 ng/g, 0.06 ± 0.01 ~ 2.50 ± 0.14 ng/g and 0.13 ± 0.10 ~ 3.64 ± 0.74 ng/g, respectively. This developed method was successfully applied to the determination of antibiotics in different tissues of hydroponic wetland plants exposed to antibiotics-spiked water for one-month. Sixteen of 28 spiked antibiotics were detected in plant tissue samples. Overall, of these 16 antibiotics, all were detected in root samples (from < LOQ to 1478 ± 353 ng/g), eleven in stem samples (from < LOQ to 425 ± 47.0 ng/g), and nine in leaf samples (from < LOQ to 429 ± 84.5 ng/g). This developed analytical method provided a robust tool for the simultaneous screening and determination of antibiotics in different plant tissues.

circAFF1 Aggravates Vascular Endothelial Cell Dysfunction Mediated by miR-516b/SAV1/YAP1 Axis.

Pathological vascular endothelial damage caused by hypoxia is the basis of many vascular-related diseases. However, the role of circular RNA in hypoxic vascular injury is still poorly understood. Here, we found that hypoxia induced AFF1 circular RNA (circAFF1) can activate the SAV1/YAP1 and lead to the dysfunction of vascular endothelial cells. In HUV-EC-C and HBEC-5i cells, circAFF1 was upregulated under CoCl2 induced hypoxic conditions. The abnormal expression of circAFF1 inhibited the proliferation, tube formation, migration of vascular endothelial cells. The effect of circAFF1 is achieved by the adsorption of miR-516b to release SAV1, which in turn causes the phosphorylation of YAP1. Moreover, we found that the upregulation of circAFF1 in 235 Patients with subarachnoid hemorrhage. Taken together, we clarify the role of circAFF1/miR-516b/SAV1/YAP1 axis in vascular endothelial dysfunction and its potential early diagnostic value of disease caused by hypoxia injury in blood vessels.