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Experimental research on flow characteristics of open natural circulation system was performed, to figure out the mechanism of the open natural circulation behaviors. The influence factors, such as the heating power, the inlet subcooled and the level of cooling tank on the flow characteristics of the system were examined. It was shown that within the scope of the experimental conditions, there are five flow types: single-phase stable flow, flash and geyser coexisting unstable flow, flash stable flow, flash unstable flow, and flash and boiling coexisting unstable flow. The geyser flow in flash and geyser coexisting unstable flow is different from classic geysers flow. The flow oscillation period and amplitude of the former are more regular, is a newly discovered flow pattern. By drawing the flow instability boundary diagram and sorting out the flow types, it is found that the two-phase unstable flow is mainly characterized by boiling and flash, which determine the behavior of open natural circulation respectively or jointly. Moreover, compared with full liquid level system, non-full liquid level system is more prone to boiling phenomenon, and the range of heat flux density and undercooling degree corresponding to unstable flow is larger.

Background: The ability to differentiate stimuli that predict fear is critical for survival; however, the underlying molecular and circuit mechanisms remain poorly understood., Methods: We combined transgenic mice, in vivo transsynaptic circuit-dissecting anatomical approaches, optogenetics, pharmacological methods, and electrophysiological recording to investigate the involvement of specific extended amygdala circuits in different fear memory., Results: We identified the projections from central lateral amygdala (CeL) protein kinase C δ (PKCδ)-positive neurons and somatostatin (SST)-positive neurons to GABAergic (gamma-aminobutyric acidergic) and glutamatergic neurons in the ventral part of the bed nucleus of stria terminalis (vBNST). Prolonged optogenetic activation or inhibition of the PKCδ CeL-vBNST pathway specifically reduced context fear memory, whereas the SST CeL-vBNST pathway mainly reduced tone fear memory. Intriguingly, optogenetic manipulation of vBNST neurons that received the projection from PKCδ CeL neurons exerted bidirectional regulation of context fear, whereas manipulation of vBNST neurons that received the projection from SST CeL neurons could bidirectionally regulate both context and tone fear memory. We subsequently demonstrated the presence of δ and κ opioid receptor protein expression within the CeL-vBNST circuits, potentially accounting for the discrepancy between prolonged activation of GABAergic circuits and inhibition of downstream vBNST neurons. Finally, administration of an opioid receptor antagonist cocktail on the PKCδ CeL-vBNST or SST CeL-vBNST pathway successfully restored context or tone fear memory reduction induced by prolonged activation of the circuits., Conclusions: Together, these findings establish a functional role for distinct CeL-vBNST circuits in the differential regulation and appropriate maintenance of fear., (Copyright © 2023 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.)

Objective: This study aimed to develop a simple, sensitive, and selective Liquid chromatography with a Mass spectroscopic method for simultaneous quantification of a recombinant bifunctional hirudin (r-RGD-Hirudin, Bifunctional Hirudin, BFH) in human plasma and verify its effectiveness., Methods: The analytes and the internal standards from human plasma were extracted using the solid-phase extraction technique. The reconstituted samples were chromatographed on Waters C 18 column (BEH 50 × 2.1 mm, 1.7 μm) using a mixture of 0.1% formic acid/acetonitrile (85%/15%, v/v) with gradient elution as the initial mobile phase at a flow rate of 0.3 mL/min., Results: The effectiveness of the proposed method was verified over the concentration range of 10-2000 ng/mL for r-RGD-Hirudin. A linear calibration curve was obtained. The precision and accuracy of BFH in the intra- and inter-day runs fell within the range of ±15% at LQC, GMQC, MQC and HQC concentrations. The extraction recoveries and matrix effect at two quality control (QC) levels for BFH were confirmed to conform to the relevant requirement., Conclusion: The proposed method was successfully adapted to examine the pharmacokinetics of BFH in 40 Chinese healthy volunteers, respectively., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Feasible and accurate predictors are urgently needed to evaluate the survival for patients with paraquat poisoning since the high mortality of paraquat poisoning always resulted in the loss of both life and money. Multiple predictors have been developed to predict prognosis of the patients with PQ poisoning, which however heavily depend on the time of admission to hospitals. Here we reported a feasible and accurate prognosis predictor for patients with paraquat poisoning that is independent of the time of admission to hospitals. Patients with paraquat poisoning were enrolled in this study according to the inclusion and exclusion criteria, which were grouped into survivors and non-survivors based on the 90-days follow-up investigation. The concentration of paraquat in serum and urine, and the baseline clinical parameters associated with the injuries of the liver, kidney, and lung were evaluated to predict the survival of these patients by using receiver operating characteristic curve (ROC) analysis, univariate and multivariate cox regression analyses. A total of 114 patients was included in this study with a survival rate of 54.4%. The median survival days of non-survivors were 6.0 (95%Cl: 4.0-7.8). A new predictor, namely paraquat concentration-associated multiorgan injury index (PCAMII), was established by integrating serum and urine paraquat concentration, serum creatinine, alanine aminotransferase, aspartate transaminase, total and direct bilirubin, at different weighting coefficients, with the accuracy of about 90%. The model to predict the survival probability by PCAMII was established with good fitness (R 2  = 0.9325), providing the simulated survival rates comparable to the clinical data. PCAMII, which is independent of hospital admission time, is a feasible and accurate marker to predict the survival rate of patients with PQ poisoning., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Millettia pulchra is a renowned anti-inflammatory herbal medicine in southeast provinces of China. However, the underlying anti-inflammation mechanism remained incompletely understood. Herein, four new isoflavones, pulvones A-D and eleven reported constituents were isolated from the stems of Millettia pulchra with their structures being elucidated by HRMS and NMR analysis. The anti-inflammatory activities of pulvones A and C were further evaluated due to the better inhibitory activity on nitric oxide production in LPS-stimulated RAW264.7 cells and no obvious cytotoxicity to RAW264.7 cells. Western blot showed that pulvones A significantly decreased the levels of iNOS and COX-2 proteins and pulvones C only decreased the level of iNOS protein. ELISA analysis demonstrated that pulvones A inhibited the production of both interleukin-6 (IL-6) and IL-1β while pulvones C showed better suppression effect on IL-1β production in LPS-stimulated RAW264.7 cells. Then, their potential inhibitory effects on NF-κB pathway were tested in LPS-stimulated RAW264.7 cells. Immunofluorescence and western blot assay showed that pulvones A and C reduced the nuclear translocation of NF-κB(p65) and interrupted IκB phosphorylation. The ADP-Glo™ kinase assay showed pulvones A and C could directedly inhibit the IKKβ kinase activity with the inhibitory rate of 40%, which were also verified by docking study. Collectively, these results suggested that pulvones A and C's anti-inflammatory effects were relevant to the interruption of NF-κB activation by inhibiting IKKβ kinase., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)

In this paper, molecular imprinting and photonic crystal techniques were combined to construct a four-channel sensor array for the simultaneous identification of various sulfonamides. The assay was composed of four units. Three of these units were prepared using sulfaguanidine, sulfamethazine, or sulfathiazole as template molecules. The fourth unit was prepared without a template molecule. The preparation was optimized to obtain maximum identification with a molar ratio of template, monomer, and cross-linker of 1:50:10. The response time was as short as 10 min. For demonstration, six sulfonamides were selected as analytes. The Bragg diffraction patterns of analytes at different concentrations were measured using the sensor array. Data obtained were analyzed using linear discrimination analysis (LDA) and principal component analysis (PCA). LDA can be applied for SAs discrimination. The message ratios of 87.6%, 94.4%, and 95.8% for six SAs at 10 -4  mol L -1 , 10 -6  mol L -1 , and 10 -8  mol L -1 were achieved using LDA. The sensor array identified the mixture containing various SAs with an LDA coefficient of 86.1%, thereby indicating that the sensor array had a strong anti-interference ability. The sensor array was used to identify six SAs in fish samples. The measured data in spiked samples were consistent with the fingerprint collected from standard solutions. The accuracy rate reached 90.9%, indicating that the array can be used to identify SAs from food samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Objective: Postoperative ileus (PI) is a common complication following posterior thoraco-lumbar spinal fusion surgery. It usually slows patient's recovery and increases postoperative morbidity. However, the risk factors associated with PI in patients undergoing posterior thoraco-lumbar spinal fusion surgery are still unclear. The purpose of this study is to investigate the potential risk factors for PI in those patients., Patients and Methods: A prospective study was conducted and 426 patients received posterior thoraco-lumbar spinal fusion surgery between March 2017 and February 2018 were included in this study. The associations between different clinical factors and PI were analyzed. A logistic regression analysis was performed to detect independent risk factors for PI. The cut-off value, sensitivity and specificity of these independent factors were calculated by receiver operating characteristic (ROC) curve., Results: In this study, 8.2% (35/426) of these patients were identified with PI. The average length of postoperative hospital stay was 12.54 ± 6.06 days in patients with PI compared with 8.91 ± 3.81 days in patients without PI (P =  0.001). These results indicated that surgical duration, PLIF approach, blood loss and length of postoperative diet restriction were potential risk factors for PI in patients with thoraco-lumbar spinal fusion surgery. The cut-off values of surgical duration, blood loss and length of postoperative diet restriction were 4.375 h, 750 ml and 9.5 h, respectively. Combination of surgical duration, PLIF approach, blood loss and length of postoperative diet restriction has the highest predictive value for PI (AUC = 0.910, P <  0.001)., Conclusion: Based on the study, surgical duration, PLIF approach, blood loss and length of postoperative diet restriction were the independent risk factors for PI in patients with posterior thoraco-lumbar spinal fusion surgery. Combined those factors has the highest risk for developing PI., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: For patients undergoing lumbar spinal surgery, many surgeons routinely perform laboratory tests within 3 days after surgery. However, few studies have reported the necessity for routine laboratory tests for patients with uncomplicated cases within 3 days after surgery., Methods: We performed a retrospective study of patients with lumbar degenerative disease who had undergone lumbar spinal surgery from May 2014 to May 2017. The perioperative patient information was recorded. The abnormal postoperative laboratory tests were recorded. Finally, the incidence and risk factors for patients requiring postoperative clinical treatment were analyzed., Results: A total of 1915 patients were included in the present study. Postoperative laboratory tests had been ordered for 870 patients (45.43%). Of these patients, only a small proportion had required postoperative clinical intervention to treat abnormal serum hemoglobin (2.53%), albumin (1.95%), serum potassium (0.92%), or serum calcium (6.55%) levels. Multivariate logistic regression analysis showed that female gender and operative time were risk factors for the need for blood transfusion after lumbar spinal surgery. Age and operative time were risk factors for patients requiring albumin supplementation after lumbar spinal surgery. Finally, intraoperative blood loss and operative time were independent risk factors for patients requiring calcium supplementation after surgery., Conclusions: Owing to the small number of postoperative clinical interventions for abnormal laboratory test results, we believe that the use of routine laboratory tests within 3 days after lumbar spinal surgery for patients with uncomplicated cases are unnecessary. Our results showed that operative time is a potential risk factor for the necessity for clinical treatment after lumbar spinal surgery., (Copyright © 2019 Elsevier Inc. All rights reserved.)

It is known that inflammation and oxidative stress have strong influences on chronic kidney disease (CKD). As an antioxidant, bilirubin is currently under extensive scrutiny. However, there are disagreements with regard to the oxidative and antioxidative roles of serum uric acid (SUA). This study aimed to investigate the relationship between serum bilirubin and the progression of renal function in CKD patients with hyperuricemia (HUA). This retrospective longitudinal study included 427 CKD patients. The endpoint was renal replacement therapy or death. Patients were divided into the following two groups according to the SUA level: HUA group (SUA ≥ 420 μmol/L for men; SUA ≥ 360 μmol/L for women) and normal uric acid level (NUA) group. A Cox proportional hazards model was used to evaluate the risk factors for renal outcomes in the two patient groups. The median follow-up time was 36 months. In the Cox regression analysis, the risk of renal outcomes in patients with serum indirect bilirubin (IBIL) levels >4.55 μmol/L was 0.15 times the risk in patients with serum IBIL levels ≤4.55 μmol/L (hazard ratio = 0.15, p = .013). Our findings suggest that a high serum IBIL level might be a protective factor for the progression of renal function in CKD patients with HUA., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: Oxidative stress and inflammation play pivotal roles in chronic kidney disease (CKD). Bilirubin is an endogenous anti-inflammatory antioxidant. However, the relationship between serum bilirubin and renal outcomes in CKD is controversial. We explored the association of serum bilirubin levels with renal outcomes in Han Chinese patients with CKD., Methods: Clinical and laboratory data were collected from 316 patients with CKD. The primary clinical endpoint was renal replacement therapy or death. The association between serum bilirubin and clinical parameters was assessed by correlation analysis. Multiple Cox regression analysis was used to explore the relationship between serum bilirubin and renal outcomes in patients with CKD., Results: Serum total and indirect bilirubin were positively correlated with estimated glomerular filtration rate, but negatively correlated with 24-h urine protein in patients with CKD. Serum total and indirect bilirubin were inversely associated with CKD stages in patients with CKD stages 1-5. Multiple Cox regression analysis demonstrated that the higher concentration of serum total bilirubin was independently associated with better renal outcomes in CKD., Conclusions: Our results suggest that serum total bilirubin may have protective effects on kidneys., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Insulin or insulin like signaling (IIS) pathway is a crucial pathway in Caenorhabditis elegans associated with mediating longevity, and stress resistance. Regulators of G protein signaling (RGS) also modulate stress resistance and longevity in multiple in vitro and in vivo models. However, the mechanism underlying RGS mediating stress resistance and longevity remains largely unclear. Here we report that rgs-1, an important member of rgs family, is a novel modulator of IIS pathway in C. elegans. We found that the loss of rgs-1 dramatically promoted paraquat resistance in C. elegans. Further genetic analyses demonstrated that rgs-1 acted downstream of daf-2 and upstream of age-1, pdk-1, daf-16. Instead of affecting those IIS-associated genes in transcriptional process, loss of rgs-1 promoted DAF-16's nucleus translocation and subset genes' expression in paraquat-induced oxidative status. By this way, rgs-1 mutant worms exhibited lower ROS damage and longer survival time than wild type worms when both exposed to paraquat. Other than paraquat exposure, rgs-1 mutant also promoted lifespan and cadmium resistance relying on daf-16. As rgs is evolutionarily conserved, our findings open a new insight into rgs family and its role in paraquat-induced oxidative stress and longevity in C. elegans or even mammals., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Objective: Anterior cervical discectomy and fusion (ACDF) is a popular procedure for patients with cervical spondylotic myelopathy, but few studies reported the clinical outcomes of cervical local bone graft with a PEEK cage used in it. This retrospective study was performed to compare the clinical and radiological outcomes of using local bone graft with a PEEK cage versus iliac bone graft in ACDF., Patients and Methods: A total of 60 consecutive patients who underwent ACDF were evaluated from January 2010 to January 2013. Twenty-nine patients received ACDF with a PEEK cage combined with cervical local bone graft (local bone group) and 31 patients received ACDF with autologous tricortical iliac bone graft (iliac bone group). The intraoperative and perioperative complications of both groups were recorded. Preoperative and postoperative radiographs were taken to calculate the ratio of interbody height to the disc height and the interbody bony fusion rate. The Japanese Orthopedic Association (JOA) score and visual analogue scale (VAS) were used to estimate postoperative clinical outcomes., Results: The mean follow-up duration was 25.0±3.8months in the local bone group and 24.4±3.4months in the iliac bone group (P=0.56). Although there was no significant difference between the two groups in terms of blood loss (P=0.17), the length of surgery was significantly less in the local bone group comparing with that of iliac bone group (P=0.01). Postoperatively, VAS scores were significantly decreased, and JOA scores were improved in both groups. However, no statistically significant differences were found between the two groups at final follow up (P=0.45 and P=0.93). The disc space height and segmental interbody angle at the surgical segment were greater in local bone group than those in the iliac bone group (P<0.001 and P<0.001). The fusion rates were 93.1% in local bone group and 90.3% in the iliac bone group at last follow up (P=0.70). Perioperative complication rates in local bone group and iliac bone groups were 6.8% and 29%, respectively (P=0.04)., Conclusions: Based on this study, patients receiving ACDF with local bone graft combined with a PEEK cage had significant shorter operation time, lower perioperative complications rate, and better radiological results comparing with those with an iliac bone graft alone. It seems that the local bone graft with a PEEK cage appears to be a safe alternative to the iliac bone graft for ACDF., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Objective: Lumbosacral spinal tuberculosis is rare in current population. Previous studies have reported effective outcomes about anterior, antero-posterior and posterior surgery for treating tuberculosis of lumbosacral region. However, the bone grafts used in these studies are mainly structural bone and mesh cage. The purpose of this study is to evaluate the efficacy and safety of nonstructural autograft in the surgical treatment of lumbosacral tuberculosis by one-stage posterior procedure., Patients and Methods: A total of 21 patients with lumbosacral tuberculosis were retrospectively reviewed between January 2012 and December 2014. All the patients underwent one-stage posterior debridement, interbody fusion with nonstructural autograft and posterior instrumentation. The preoperative and postoperative erythrocyte sedimentation rates (ESR), C-reactive protein (CRP) and visual analogue scale (VAS) were recorded. Preoperative and postoperative lumbosacral angle and intervertebral space height were measured on the plain films. American Spinal Injury Association (ASIA) Impairment Scale was used to evaluate the neurological outcomes of the patients., Results: The average follow up period was 22.9±6.7months (range 12-36 months). The preoperative ESR and CRP were 33.4±10.5mm/h and 30.3±20.3mg/l, respectively, which decreased to 15.2±7.1mm/h and 10.6±5.8mg/l postoperatively with significant differences (P<0.05). The lumbosacral angles and intervertebral space height were increased from preoperative 20.4°±4.5° and 9.7±1.9mm to postoperative 25.6°±4.6° and 12.3±2.1mm, respectively (P<0.001 and P<0.001). At the final follow up, a loss of 2.1°of lumbosacral angles and 1.6mm of intervertebral space height was observed. The VAS scores were decreased from 4.73 to 2.71. Bony fusion was achieved in all patients at 6 months after surgery. Neurological outcomes were improved with 1-2 grades in most of the patients. One patient got wound infection and was cured by daily dressing. Complications related to instrumentation or neurological deficit weren't observed., Conclusion: Combined with one-stage posterior debridement and instrumentation, interbody fusion with nonstructural autograft is an effective option for lumbosacral tuberculosis., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A direct competitive enzyme-linked immunosorbent assay (ELISA) method was used for the detection of malachite green (MG) with a high sensitivity and selectivity using magnetic molecularly imprinted polymers (MMIPs) as a bionic antibody. MMIPs were prepared through emulsion polymerization using Fe 3 O 4 nanoparticles as magnetic nuclei, MG as a template, methacrylic acid (MAA) as a functional monomer, ethylene glycol dimethacrylate (EGDMA) as a crosslinking agent and span-80/tween-80 as mixed emulsifiers. The MMIPs were characterized by scanning electron micrographs (SEM), thermal-gravimetric analyzer (TGA), Fourier transform infrared spectrometer (FT-IR) and vibrating sample magnetometer (VSM), respectively. A high magnetic saturation value of 54.1emug -1 was obtained, resulting in rapid magnetic separation of MMIPs with an external magnet. The IC 50 of the established ELISA method was 20.1μgL -1 and the detection limit (based on IC 85 ) was 0.1μgL -1 . The MMIPs exhibited high selective binding capacity for MG with cross-reactivities less than 3.9% for MG structural analogues. The MG spiking recoveries were 85.0%-106% with the relative standard deviations less than 4.7%. The results showed that the biomimetic ELISA method by using MMIPs as bionic antibody could be used to detect MG rapidly in fish samples with a high sensitivity and accuracy., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Objective: With the growing need for reliable and durable upper arm hemodialysis access, we sought to compare the performance of a novel modified nontransposed brachiobasilic arteriovenous fistula (mNT-BBAVF) with that of the more traditional brachiocephalic arteriovenous fistula (BCAVF)., Methods: Briefly, to construct an mNT-BBAVF, an incision is made on the ulnar side of the elbow. The brachial artery and basilic vein are then isolated, and a side-to-side anastomosis is performed without transposition of the basilic vein. Next, the proximal basilic vein and the perforating veins within the surgical field are ligated. In this study, we retrospectively reviewed the medical records of all patients who underwent either an mNT-BBAVF or a BCAVF between January 2011 and October 2014 to compare 1-year primary unassisted patency, cumulative patency, and complications. We also examined hemodynamic parameters of vessels in each fistula type., Results: We identified a total of 84 patients: 45 had a BCAVF, and 39 had an mNT-BBAVF. The two groups were well matched for baseline characteristics. Maturation rates at 1 month were 97% for mNT-BBAVF and 96% for BCAVF. The 1-year primary unassisted patency was significantly higher in the mNT-BBAVF group than that in the BCAVF group (87% vs 67%; hazard ratio, 2.86; 95% confidence interval, 1.11-6.40; P = .03), although cumulative patency did not differ (90% vs 73%; hazard ratio, 2.80; 95% confidence interval, 0.98-6.96 ; P = .06). There were no differences in thrombosis, failure of maturation, bleeding, steal syndrome, arm swelling, aneurysm, and stenosis between the two groups during the 12-month study. Importantly, diameters and blood flow volumes of the proximal cephalic vein, distal cephalic vein, and distal basilic vein in patients who received an mNT-BBAVF increased significantly after 12 months. All three vessels met the Kidney Disease Outcomes Quality Initiative (KDOQI) criteria for fistula maturation and were available for dialysis cannulation, whereas only the proximal cephalic vein in the BCAVF group met the maturation criteria and could be used for cannulation., Conclusions: mNT-BBAVF appeared to be an effective alternative to BCAVF for upper arm hemodialysis access., (Copyright © 2016 Society for Vascular Surgery. All rights reserved.)

Aims: Paraquat (PQ) is a pesticide highly toxic to human beings, and a well-known trigger of oxidative stress. Although several animal models of PQ poisoning have been developed, some disadvantages limit their application in vivo. A zebrafish model was used in the present study to better define mechanisms of oxidative stress injury induced by PQ., Main Methods: The toxicity of PQ was evaluated in the AB strain of zebrafish, and apoptosis was assessed by acridine orange staining. Macrophage migration was identified using the TG (zlyz:EGFP) transgenic strain, and angiogenesis was observed using the fli1a-EGFP casper strain. Following the validation of gene changes by zebrafish-based in vivo quantitative real-time PCR, network analysis was performed using the Ingenuity Pathway Analysis software., Key Findings: We first established the LC50 of PQ in the zebrafish model, and then found that robust oxidative stress and antioxidant genes were activated after PQ exposure. Moreover, apoptosis and distinct macrophage activation and migration were identified for the first time in PQ-exposed zebrafish. Utilizing this model, both extrinsic and intrinsic pathways involved in PQ-induced apoptosis were elucidated. We further demonstrated that macrophage migration was specifically induced by PQ, and that Rho family members and JNK-MMP13 signaling participated in this process., Significance: Zebrafish is a promising tool for investigating the mechanisms of oxidative stress injury induced by PQ, and for screening effective anti-oxidant drugs., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Aims: Hyperphosphatemia is associated with accelerated vascular endothelial dysfunction in patients with chronic kidney disease (CKD). The purpose of this study is to investigate the molecular mechanisms underlying hyperphosphatemia-caused endothelial dysfunction., Main Methods: The metabolic fingerprinting of human umbilical vein endothelial cells (HUVECs) subjected to hyperphosphatemia was characterized using an integrated metabolomics approach. HUVECs cultured in physiologically simulated hyperphosphatemia with or without phosphonoformic acid, a sodium-dependent phosphate transporter inhibitor (N=6) were collected for metabolomics analysis. Multivariate principle component analysis and partial least squares discriminant analysis were applied to analyze the metabolic data. The key metabolites were confirmed by quantitative analysis using liquid chromatography coupled with tandem mass spectrometer (LC-MS/MS)., Key Findings: 36 metabolites were significantly altered in HUVECs following the challenges of hyperphosphatemia mimic, involving several metabolic pathways (all P<0.05). Among them, ornithine increased significantly in the HUVECs mediated by hyperphosphatemia mimic, and its levels positively correlated with cell apoptosis rate (r=0.674, P=0.002), and several additional metabolites in multiple metabolic pathways. The changes in the levels of ornithine and other several metabolites were supported by subsequent quantitative analyses using LC-MS/MS. Further study demonstrated that the increase in ornithine level may result from the increased expression of arginase 2 in HUVECs, which mediates the hydrolysis of arginine to form ornithine., Significance: This is the first study demonstrating ornithine a key molecule mediating hyperphosphatemia-induced apoptosis of ECs. Arginase 2 may be a therapeutic target for hyperphosphatemia-associated cardiovascular events., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Aims: Hyperphosphatemia is an independent risk factor of cardiovascular events in the patients with chronic kidney disease. High phosphate can induce endothelial cell apoptosis, but the exact mechanism is not clear. This study fills this knowledge gap., Materials and Methods: Microarray analysis was used to identify differentially expressed gene profiles in human umbilical vein endothelial cells (HUVECs) in high phosphate (3.0mM) and normal phosphate (1.0mM) medium. Microarray informatics analysis was used to explore key pathways and genes. High phosphate-induced apoptosis is marked by annexin V-FITC/PI staining and cleavage of caspase-3. Immunoblotting and quantitative real-time PCR were performed to identify the microarray analysis., Key Findings: Our microarray informatics analysis reveals that the mitogen-activated protein kinase (MAPK) plays a key role. As suggested by gene coexpression network analysis, bone morphogenetic protein 4 (BMP4) gene is a potential key regulatory gene in high phosphate environment. Both the expressions of BMP4 protein and mRNA are decreased. Extracellular regulated protein kinases (ERKs) are activated, while the inhibition of ERK by U0126 increases the expression of BMP4. Both recombinant BMP4 protein pretreatment and U0126 pretreatment reduce the apoptosis of endothelial cells in simulated hyperphosphatemia. However, BMP4 protein pretreatment had no effect on the activation of ERK MAPK pathway., Significance: Our results indicate that the inhibition of ERK MAPK pathway protects endothelial cells from apoptosis by upregulating bone morphogenetic protein 4 in endothelial cells exposed to hyperphosphatemia. Our study provides potential molecular targets for developing new strategies to reduce the endothelial cell apoptosis induced by high phosphate., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) is regulated by a myriad of signaling cascades including glycogen synthase kinase (GSK) 3β and plays a Janus role in podocyte injury. In vitro, lipopolysaccharide (LPS) or adriamycin (ADR) elicited podocyte injury and cytoskeletal disruption, associated with NFκB activation and induced expression of NFκB target molecules, including pro-survival Bcl-xL and podocytopathic mediators like MCP-1, cathepsin L, and B7-1. Broad-range inhibition of NFκB diminished the expression of all NFκB target genes, restored cytoskeleton integrity, but potentiated apoptosis. In contrast, blockade of GSK3β by lithium or 4-benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione (TDZD-8) mitigated the expression of podocytopathic mediators, ameliorated podocyte injury, but barely affected Bcl-xL expression or sensitized apoptosis. Mechanistically, GSK3β was sufficient and essential for RelA/p65 phosphorylation, specifically at serine 467, which specifies the expression of selective NFκB target molecules, including podocytopathic mediators, but not Bcl-xL. In vivo, lithium or TDZD-8 therapy improved podocyte injury and proteinuria in mice treated with LPS or ADR, concomitant with the suppression of podocytopathic mediators, but retained Bcl-xL in glomerulus. Broad-range inhibition of NFκB conferred similar but much weakened antiproteinuric and podoprotective effects accompanied with a blunted glomerular expression of Bcl-xL and marked podocyte apoptosis. Thus, the GSK3β-dictated fine-tuning of NFκB may serve as a novel therapeutic target for podocytopathy.

Aims: Paraquat Poisoning (PQ) can cause illness and death, and its main causes of mortality are acute respiratory failure and lung fibrosis. Early recognition of this condition and early treatment are vital. Thus, it is of importance to target the key genes controlling pathogenesis in the early stage of PQ., Main Methods: C57BL/6 mice were used for Paraquat intragastric administration as a model of PQ. Following a gene chip-based screening, the change of gene expression in the lung was further validated by bioinformatic analyses, co-expression network construction and real-time RT-PCR, Western blot and immunofluorescence assays., Key Findings: 2287 genes with differential expression were identified at the very early stage of PQ. From these, 76 genes that were linked to mitochondrion function were further pursued. Among these genes, PSTK was a phosphorylase kinase which serves a protective role in oxidative stress lung damage. PSTK was the central gene in a 30-gene network that is important for mitochondrial complex I assembly, mitochondrial apoptosis and mitochondrial fatty acid beta-oxidation, suggesting that they could conceivably be related to the pathogenesis of PQ induced lung damage. Lastly, we confirmed that PSTK was lowered in rodent lungs following PQ., Significance: PSTK emerges as a central gene in a network of mitochondrial function genes in PQ exposed mice. The functional role of PSTK in PQ induced lung injury warrants further examination., (Copyright © 2015 Elsevier Inc. All rights reserved.)

FASN plays an important role in the malignant phenotype of various tumors. Our previous studies show that inhibition FASN could induce apoptosis and inhibit proliferation in human osteosarcoma (OS) cell in vivo and vitro. The aim in this study was to investigate the effect of inhibition FASN on the activity of HER2/PI3K/AKT axis and invasion and migration of OS cell. The expression of FASN, HER2 and p-HER2(Y1248) proteins was detected by immunohistochemistry in OS tissues from 24 patients with pulmonary metastatic disease, and the relationship between FASN and p-HER2 as well as HER2 was investigated. The results showed that there was a positive correlation between FASN and HER2 as well as p-HER2 protein expression. The U-2 OS cells were transfected with either the FASN specific RNAi plasmid or the negative control RNAi plasmid. FASN mRNA was measured by RT-PCR. Western blot assays was performed to examine the protein expression of FASN, HER2, p-HER2(Y1248), PI3K, Akt and p-Akt (Ser473). Migration and invasion of cells were investigated by wound healing and transwell invasion assays. The results showed that the activity of HER2/PI3K/AKT signaling pathway was suppressed by inhibiting FASN. Meanwhile, the U-2OS cells migration and invasion were also impaired by inhibiting the activity of FASN/HER2/PI3K/AKT. Our results indicated that inhibition of FASN suppresses OS cell invasion and migration via down-regulation of the "HER2/PI3K/AKT" axis in vitro. FASN blocker may be a new therapeutic strategy in OS management., (Copyright © 2013 Elsevier Inc. All rights reserved.)

We have recently synthesized a series of phosphorylated flavonoids and identified some of them as potent inhibitors of pancreatic cholesterol esterase (CEase) with excellent selectivity for CEase over acetylcholinesterase (AChE). In the present paper, we investigated the inhibitory activities of these compounds against porcine liver carboxylesterase (CE) since carboxylesterases (CEs) are another family of serine esterases responsible for the metabolism and detoxification of many ester-containing xenobiotics and clinical esterified drugs, and there exists much structural similarity between CEase and CEs. The results indicated that phosphorylated flavonoids exhibited significantly improved inhibition potency toward CE than their parent compounds, and six of them had IC50 values less than 5.0nM. Among all compounds tested, compounds 3d and 3e are the two most potent inhibitors of CE, giving IC50 values of 1.79nM and 1.58nM, respectively. Interestingly, these compounds inhibited CEase and CE with similar structure activity correlations, and those with high inhibitory activities toward CEase could also inhibit CE efficiently. The presences of a free hydroxyl group at position 5 and a phosphate group at position 7 of the phosphorylated flavonoids are favorable to the inhibition of CE. The inhibition mechanism and kinetic characterization studies of the most potent inhibitors revealed that they are irreversible competitive inhibitors., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

The unchecked overproduction of reactive oxygen and nitrogen species by inflammatory cells can cause tissue damage, intensify inflammation, promote apoptosis, and accelerate the progression of immune-mediated glomerulonephritis (GN). Here we tested whether the anti-inflammatory and antioxidant properties of the green tea polyphenol (-)-epigallocatechin-3-gallate (EGCG) favorably affect the development of immune-mediated GN. Pretreatment of 129/svJ mice with EGCG from 2 days before to 2 weeks after the induction of GN led to reduced proteinuria and serum creatinine, and marked improvement in renal histology when compared with vehicle-pretreated diseased mice. This pretreatment reduced oxidative stress, and normalized osteopontin, p65/nuclear factor-κB, inducible nitric oxide synthase, nitric oxide metabolites, p-Akt, phosphorylated extracellular signal-regulated kinases 1 and 2, p47phox, and myeloperoxidase, all of which were elevated in vehicle-pretreated diseased mice. Levels of glutathione peroxidase and peroxisome proliferator-activated receptor-γ (PPARγ), both reduced in the vehicle-pretreated diseased mice, were normalized. This renoprotective effect was reversed by concomitant administration of the PPARγ antagonist GW9662 throughout the EGCG pretreatment period. Importantly, mortality and renal dysfunction were significantly attenuated even when the polyphenol treatment was initiated 1 week after the onset of GN. Thus, EGCG reversed the progression of immune-mediated GN in mice by targeting redox and inflammatory pathways.