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Alcohol abuse is one of the major public health problems in the world and is associated with various health conditions. However, little is known about the effect of alcohol consumption on acute kidney injury (AKI). In this study, we demonstrate that chronic and binge alcohol feeding with a Lieber-DeCarli diet containing 5 % ethanol for 10 days, followed by a single dose of 31.5 % ethanol by gavage, aggravated AKI after ischemia-reperfusion injury (IRI) in female, but not in male, mice. Kidney dysfunction, histopathology and tubular cell apoptosis were more severe in EtOH-fed female mice after IRI, compared to pair-fed controls. RNA sequencing and experimental validation uncovered that activation of integrin β1 and its downstream c-Jun NH2-terminal kinase (JNK) aggravated AKI in EtOH-fed mice. Knockdown of integrin β1 inhibited JNK phosphorylation and alleviated AKI in EtOH-fed mice, whereas activation of integrin β1 by agonist antibody increased JNK phosphorylation, worsened renal histological injury and tubular cell apoptosis, and aggravated kidney dysfunction. In vitro, activation of integrin β1 increased JNK phosphorylation and induced tubular epithelial cell apoptosis. The detrimental effect of EtOH feeding was primarily mediated by acetaldehyde, as its levels were increased in the blood, liver and kidney of female mice fed with ethanol. Acetaldehyde per se activated integrin β1/JNK signaling and induced tubular cell apoptosis in vitro. These findings suggest that alcohol consumption increases vulnerability to AKI in female mice, which is probably mediated by acetaldehyde/integrin β1/JNK signaling cascade.

Fatty acids in oat flour have been analyzed by using multiple reaction monitoring mode of gas chromatograph-mass spectrometer. The samples and fatty acid standards were methylated by potassium hydroxide - methanol and boron trifluoride- methanol solutions for 15 min to produce the corresponding fatty acid methyl ester. Then, the products would be separated through DB-FastFAME (90 m×0.25 mm×0.25 μm) capillary column. Multiple reaction monitoring mode (MRM) was performed under electron bombardment ionization (EI) source, and the external standard method was carried out for quantitative analysis. 40 kinds of fatty acid methyl esters could be completely separated within 38 min. A good linear relationship was achieved in the concentration range of 0.4~40 μg/mL. The correlation coefficients of standard curve were all greater than 0.999 7. The limit of quantification of 6 fatty acids such as C20:2 was 0.003 g/100 g, and the remaining 34 fatty acids was 0.000 6 g/100 g. The standard recoveries of fatty acids were 86.6%~107.3%, and the precision was between 0.9% and 5.2%. It is a method for simple operate and high selectivity, which has a good accurate, precise and sensitive. The method is suitable for the rapid and accurate determination of fatty acids in oat flour.

Objective To analyze the clinical features and prognostic factors of herpes simplex virus type 1 (HSV-1) encephalitis. Methods A total of 51 patients with HSV-1 encephalitis hospitalized in He'nan Provincial People's Hospital from January 2018 to September 2023 were selected, and their clinical data, laboratory and imaging examination results were recorded. The modified Rankin Scale (mRS) was used to evaluate the prognosis of patients and the recovery of neurological function after brain injury. Univariate and multivariate stepwise Logistic regression analyses were screened for risk factors of adverse prognosis of HSV-1 encephalitis. Results The main symptoms of 51 cases were fever and headache. The serological examination results showed increased white blood cell count (14 cases), increased neutrophil count (14 cases), decreased lymphocyte count (8 cases), and decreased serum sodium level (17 cases). Cerebrospinal fluid (CSF) examination by lumbar puncture showed increased CSF pressure (18 cases), increased white blood cell count (28 cases), increased protein (32 cases), and increased glucose (15 cases). There were 48 patients who underwent head MRI examination, and 43 patients (89.58%) showed obvious brain region involvement on head MRI, among which 35 patients (72.92%) with positive MRI showed typical unilateral or bilateral temporal and frontal lobe involvement, and 8 cases (16.67%) with increased cerebral pia mater enhancement. 66.67% (34/51) of the patients recovered completely within 6 months after discharge, 21.57% (11/51) of the patients had sequelae, and the fatality rate was 11.76% (6/51). There were 34 (66.67%) patients with good prognosis and 17 (33.33%) patients with adverse prognosis. Multivariate Logistic regression analysis showed age increased (OR = 1.090, 95%CI: 1.020-1.200; P = 0.023) and increased CSF glucose level (OR = 23.800, 95%CI: 23.800-1250.000; P = 0.030) were risk factors for adverse prognosis in HSV-1 encephalitis patients. Conclusions Increased aging and CSF glucose level are risk factors for the adverse prognosis in HSV - 1 encephalitis patients. In clinical practice, it is necessary to master the clinical characteristics of patients with HSV-1 encephalitis, so as to achieve early detection, diagnosis and treatment.

Introduction: The severe cases of COVID-19, a disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), often present with acute kidney injury (AKI). Although old age and preexisting medical conditions have been identified as principal risk factors for COVID-19-associated AKI, the molecular basis behind such a connection remains unknown. In this study, we investigated the pathogenic role of Klotho deficiency in COVID-19-associated AKI and explored the therapeutic potential of Klotho-derived peptide 1 (KP1).Methods: We assessed the susceptibility of Klotho deficient Kl/Kl mice to developing AKI after expression of SARS-CoV-2 N protein. The role of KP1 in ameliorating tubular injury was investigated by using cultured proximal tubular cells (HK-2) in vitro and mouse model of ischemia-reperfusion injury (IRI) in vivo.Results: Renal Klotho expression was markedly downregulated in various chronic kidney disease (CKD) models and in aged mice. Compared to wild-type counterparts, mutant KL/KL mice were susceptible to overexpression of SARS-CoV-2 N protein and developed kidney lesions resembling AKI. In vitro, expression of N protein alone induced HK-2 cells to express markers of tubular injury, cellular senescence, apoptosis and epithelial-mesenchymal transition, whereas both KP1 and Klotho abolished these lesions. Furthermore, KP1 mitigated kidney dysfunction, alleviated tubular injury and inhibited apoptosis in AKI model induced by IRI and N protein.Conclusion: These findings suggest that Klotho deficiency is a key determinant of developing COVID-19-associated AKI. As such, KP1, a small peptide recapitulating Klotho function, could be an effective therapeutic for alleviating AKI in COVID-19 patients.

Kidney fibrosis is associated with tubular injury, oxidative stress and activation of interstitial fibroblasts. However, whether these events are somehow connected is poorly understood. In this study, we show that glutathione peroxidase-3 (GPX3) depletion in renal tubular epithelium after kidney injury plays a central role in orchestrating an oxidatively stressed extracellular microenvironment, which drives interstitial fibroblast activation and proliferation. Through transcriptional profiling by RNA-sequencing, we found that the expression of GPX3 was down-regulated in various models of chronic kidney disease (CKD), which was correlated with induction of nicotinamide adenine dinucleotide phosphate (NAPDH) oxidase-4 (NOX4). By using decellularized extracellular matrix (ECM) scaffold, we demonstrated that GPX3-depleted extracellular microenvironment spontaneously induced NOX4 expression and reactive oxygen species (ROS) production in renal fibroblasts and triggered their activation and proliferation. Activation of NOX4 by advanced oxidation protein products (AOPPs) mimicked the loss of GPX3, increased the production of ROS, stimulated fibroblast activation and proliferation, and activated protein kinase C-α (PKCα)/mitogen-activated protein kinase (MAPK)/signal transducer and activator of transcription 3 (STAT3) signaling. Silencing NOX4 or inhibition of MAPK with small molecule inhibitors hampered fibroblast activation and proliferation. In mouse model of CKD, knockdown of NOX4 repressed renal fibroblast activation and proliferation and alleviated kidney fibrosis. These results indicate that loss of GPX3 orchestrates an oxidatively stressed extracellular microenvironment, which promotes fibroblast activation and proliferation through a cascade of signal transduction. Our studies underscore the crucial role of extracellular microenvironment in driving fibroblast activation and kidney fibrosis.

Wnt/β-catenin is a developmental signaling pathway that plays a crucial role in driving kidney fibrosis after injury. Activation of β-catenin is presumed to be regulated through the posttranslational protein modification. Little is known about whether β-catenin is also subjected to regulation at the posttranscriptional mRNA level. Here, we report that insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3) plays a pivotal role in regulating β-catenin. IGF2BP3 was upregulated in renal tubular epithelium of various animal models and patients with chronic kidney disease. IGF2BP3 not only was a direct downstream target of Wnt/β-catenin but also was obligatory for transducing Wnt signal. In vitro, overexpression of IGF2BP3 in kidney tubular cells induced fibrotic responses, whereas knockdown of endogenous IGF2BP3 prevented the expression of injury and fibrosis markers in tubular cells after Wnt3a stimulation. In vivo, exogenous IGF2BP3 promoted β-catenin activation and aggravated kidney fibrosis, while knockdown of IGF2BP3 ameliorated renal fibrotic lesions after obstructive injury. RNA immunoprecipitation and mRNA stability assays revealed that IGF2BP3 directly bound to β-catenin mRNA and stabilized it against degradation. Furthermore, knockdown of IGF2BP3 in tubular cells accelerated β-catenin mRNA degradation in vitro. These studies demonstrate that IGF2BP3 promotes β-catenin signaling and drives kidney fibrosis, which may be mediated through stabilizing β-catenin mRNA. Our findings uncover a previously underappreciated dimension of the complex regulation of Wnt/β-catenin signaling and suggest a potential target for therapeutic intervention of fibrotic kidney diseases.

Abstract Similar to sampling perfection with application-optimized contrast using different flip angle evolutions (SPACE), T2-weighted fast field echo (FFE) also has a black blood effect and a high imaging efficiency. The purpose of this study was to optimize 3D_T2_FFE and compare it with 3D_T2_SPACE for carotid imaging. The scanning parameter of 3D_T2_FFE was optimized for the imaging of the carotid wall. Twenty healthy volunteers and 10 patients with carotid plaque underwent cervical 3D_T2_FFE and 3D_T2_SPACE examinations. The signal-to-noise ratios of the carotid wall (SNRwall) and lumen (SNRlumen), and the contrast-to-noise ratios between the wall and lumen (CNRwall_lumen) were compared. The incidence of the residual flow signal at the carotid bifurcation and the grades of flow voids in the cerebellopontine angle region in the two sequences were also compared. The reproducibility of the two sequences was tested. No significant difference was observed between the two sequences in terms of the SNRwall of healthy individuals and patients (P = 0.132 and 0.102, respectively). The SNRlumen in the 3D_T2_FFE images was lower than that in the 3D_T2_SPACE images. No significant difference was observed between the two sequences in terms of the CNRwall-lumen. The incidence of the residual flow signal at the carotid bifurcation in 3D_T2_FFE was significantly lower than that in 3D_T2_SPACE. The grades of flow suppression in the cerebellopontine angle region in 3D_T2_SPACE was lower than that in 3D_T2_FFE. Both sequences showed excellent inter-and intra-observer reproducibility. Compared to 3D_T2_SPACE, 3D_T2_FFE showed stronger flow suppression while maintaining good imaging quality, which can be used as an alternative tool for carotid imaging.

Abstract Matrix metalloproteinase-10 (MMP-10) is a zinc-dependent endopeptidase involved in regulating a wide range of biologic processes, such as apoptosis, cell proliferation, and tissue remodeling. However, the role of MMP-10 in the pathogenesis of acute kidney injury (AKI) is unknown. In this study, we show that MMP-10 was upregulated in the kidneys and predominantly localized in the tubular epithelium in various models of AKI induced by ischemia/reperfusion (IR) or cisplatin. Overexpression of exogenous MMP-10 ameliorated AKI, manifested by decreased serum creatinine, blood urea nitrogen, tubular injury and apoptosis, and increased tubular regeneration. Conversely, knockdown of endogenous MMP-10 expression aggravated kidney injury. Interestingly, alleviation of AKI by MMP-10 in vivo was associated with the activation of epidermal growth factor receptor (EGFR) and its downstream AKT and extracellular signal-regulated kinase-1 and 2 (ERK1/2) signaling. Blockade of EGFR signaling by erlotinib abolished the MMP-10-mediated renal protection after AKI. In vitro, MMP-10 potentiated EGFR activation and protected kidney tubular cells against apoptosis induced by hypoxia/reoxygenation or cisplatin. MMP-10 was colocalized with heparin-binding EGF-like growth factor (HB-EGF) in vivo and activated it by a process of proteolytical cleavage in vitro. These studies identify HB-EGF as a previously unrecognized substrate of MMP-10. Our findings also underscore that MMP-10 can protect against AKI by augmenting EGFR signaling, leading to promotion of tubular cell survival and proliferation after injury.

Objective To investigate the mechanism of cigarette smoke extract (CSE) inducing apoptosis of pulmonary artery endothelial cells (HPAECs), especially focus on the role of NLRP3(NLR family pyrin domain protein 3) inflammasome. Methods After transfection with NLRP3 siRNA and NC siRNA, HPAECs from experimental group and control group were stimulated by 10% CSE for 12 h, annexin V/PI flow method was used to detect the apoptosis of the two groups of cells before and after CSE stimulation. Cultured HPAECs were further divided into four groups:control group, CSE group (10% CSE solution was added), NAC+CSE group (10 mmol/L NAC pretreatment for 1 h and then 10% CSE culture solution) and NAC group (10 mmol/L NAC treatment for 1 h). After 12 h culture, intracellular reactive oxygen species(ROS) were detected by DCFH-DA fluorescent probe. The expression of NLRP3 and caspase-1 was determined by Western blot. Results After CSE stimulation, the survival rate of HPAECs in siNC group decreased, and the apoptosis was significantly increased as compared with that before CSE stimulation (P＜0.05). After CSE stimulation, the apoptosis rate of siNLRP3 group decreased compared with that after the same CSE stimulation in siNC group (P＜0.05).The intracellular ROS level in the CSE group was significantly increased as compared with that in the control group, which in the NAC+CSE group was decreased compared with that in the CSE group (P＜0.05). In the CSE group, NLRP3 and caspase-1 were significantly increased compared with the control group, while the expression of NLRP3 and caspase-1 in NAC+CSE group was lower than that in CSE group (P＜0.05). Conclusions CSE could induce apoptosis of HPAECs accompanied by increased NLRP3 inflammatory corpuscle expression. Silencing of NLRP3 protein expression can inhibit the apoptotic effect of CSE on HPAEC.

Podocyte injury and proteinuria are the most common features of glomerular disease, which is the leading cause of end-stage renal failure. Hyperactivated Wnt/β-catenin signaling is closely associated with podocyte injury, but the underlying mechanisms are incompletely understood. Here we show that miRNA-671-5p (miR-671-5p) plays a crucial role in mediating β-catenin-triggered podocyte injury by targeting Wilms tumor 1 (WT1). Microarray-based expression profiling revealed that miR-671-5p was the most upregulated miRNA in podocytes after β-catenin activation. MiR-671-5p was colocalized with β-catenin in the glomeruli of proteinuric CKD in vivo. Bioinformatics analyses and luciferase reporter assays confirmed that miR-671-5p targeted WT1 mRNA. Overexpression of miR-671-5p mimics inhibited WT1 and impaired podocyte integrity, whereas miR-671-5p antagomir preserved the expression of WT1 and other podocyte-specific proteins under basal conditions or after β-catenin activation. In mouse remnant kidney model, overexpression of miR-671-5p aggravated podocyte injury, worsened kidney dysfunction and exacerbated renal fibrosis after 5/6 nephrectomy. In contrast, miR-671-5p antagomir alleviated podocyte injury and attenuated proteinuria and renal fibrotic lesions after glomerular injury in vivo. These studies underscore a pivotal role of miR-671-5p in mediating WT1 depletion and podocyte injury induced by β-catenin. Targeting miR-671-5p may serve as a new approach to prevent podocyte injury and proteinuria in proteinuric CKD.

Studies suggest that Wnt/β-catenin agonists are beneficial in the treatment of acute kidney injury (AKI); however, it remains elusive about its role in the prevention of AKI and its progression to chronic kidney disease (CKD). In this study, renal Wnt/β-catenin signaling was either activated by overexpression of exogenous Wnt1 or inhibited by administration with ICG-001, a small molecule inhibitor of β-catenin signaling, before mice were subjected to ischemia/reperfusion injury (IRI) to induce AKI and subsequent CKD. Our results showed that in vivo expression of exogenous Wnt1 before IR protected mice against AKI, and impeded the progression of AKI to CKD in mice, as evidenced by both blood biochemical and kidney histological analyses. In contrast, pre-treatment of ICG-001 before IR had no effect on renal Wnt/β-catenin signaling or the progression of AKI to CKD. Mechanistically, in vivo expression of exogenous Wnt1 before IR suppressed the expression of proapoptotic proteins in AKI mice, and reduced inflammatory responses in both AKI and CKD mice. Additionally, exogenous Wnt1 inhibited apoptosis of tubular cells induced by hypoxia-reoxygenation (H/R) treatment in vitro. To conclude, the present study provides evidences to support the preventive effect of Wnt/β-catenin activation on IR-related AKI and its subsequent progression to CKD.

The effects of milling process parameters on cutting forces and surface roughness were studied during peripheral up milling of bamboo scrimber. The study results indicated that the effect of feed rate on cutting force components Fx, +Fy, -Fy, and surface roughness Ra were the most significant compared to spindle speed and cut depth during longitudinal milling. Fx, +Fy, -Fy, and Ra decreased slightly with increasing spindle speed, but increased greatly with increasing feed rate. In addition, cutting direction had a great effect on the cutting force components Fx and +Fy, as well as the machined surface roughness Ra. Fx, +Fy, and Ra during longitudinal cutting were always larger than those during end-grain cutting. Decreasing the feed rate appropriately could greatly improve the quality of the machined surface.