Asymmetric Construction of Carbon-Fluorine Quaternary Stereogenic Centers via Synergistic Pd/Cu Catalysis.

We developed an asymmetric decarboxylative allylic alkylation of vinylethylene carbonates with α-fluoro pyridinyl acetates through a synergistic palladium/copper catalysis. This protocol provides chiral allylic alcohol with carbon-fluorine quaternary stereogenic centers in good yield with good enantioselectivities and excellent regioselectivities. The utility of this approach was further demonstrated via a gram-scale experiment and derivatizations of the product.

Exploration of Damage Identification Method for a Large-Span Timber Lattice Shell Structure in Taiyuan Botanical Garden based on Structural Health Monitoring.

Large-span spatial lattice structures generally have characteristics such as incomplete modal information, high modal density, and high degrees of freedom. To address the problem of misjudgment in the damage detection of large-span spatial structures caused by these characteristics, this paper proposed a damage identification method based on time series models. Firstly, the order of the autoregressive moving average (ARMA) model was selected based on the Akaike information criterion (AIC). Then, the long autoregressive method was used to estimate the parameters of the ARMA model and extract the residual sequence of the autocorrelation part of the model. Furthermore, principal component analysis (PCA) was introduced to reduce the dimensionality of the model while retaining the characteristic values. Finally, the Mahalanobis distance (MD) was used to construct the damage sensitive feature (DSF). The dome of Taiyuan Botanical Garden in China is one of the largest non-triangular timber lattice shells worldwide. Relying on the structural health monitoring (SHM) project of this structure, this paper verified the effectiveness of the damage identification model through numerical simulation and determined the damage degree of the dome structure through SHM measurement data. The results demonstrated that the proposed damage identification method can effectively identify the damage of large-span timber lattice structures, locate the damage position, and estimate the degree of damage. The constructed DSF had relatively strong robustness to small damage and environmental noise and has practical application value for SHM in engineering.

Experimental Study on Variable-Amplitude Fatigue Performance of M60 High-Strength Bolts of Grid Structures with Bolted Spherical Joints.

The high-strength bolts of grid structures with bolted spherical joints under the action of suspension cranes are at risk of severe fatigue failure. Thus, this paper studies the variable-amplitude fatigue performance of M60 high-strength bolts. The test results for eight specimens in four loading modes are obtained using an Amsler fatigue testing machine. The fatigue life is also estimated based on Miner and Corten-Dolan's theories, and the applicability of Corten-Dolan's theory is verified. The fracture induced by the variable-amplitude fatigue is microscopically analyzed using scanning electron microscopy (SEM), revealing the mechanism of the variable-amplitude fatigue failure. Our findings provide valuable experimental data supporting the fatigue life estimation of grid structures with bolted spherical joints in service.

KIF26B-AS1 Regulates TLR4 and Activates the TLR4 Signaling Pathway to Promote Malignant Progression of Laryngeal Cancer.

Laryngeal cancer is one of the highest incidence, most prevalently diagnosed head and neck cancers, making it critically necessary to probe effective targets for laryngeal cancer treatment. Here, real-time quantitative reverse transcription PCR (qRT-PCR) and western blot analysis were used to detect gene expression levels in laryngeal cancer cell lines. Fluorescence in situ hybridization (FISH) and subcellular fractionation assays were used to detect the subcellular location. Functional assays encompassing Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), transwell and wound healing assays were performed to examine the effects of target genes on cell proliferation and migration in laryngeal cancer. The in vivo effects were proved by animal experiments. RNA-binding protein immunoprecipitation (RIP), RNA pulldown and luciferase reporter assays were used to investigate the underlying regulatory mechanisms. The results showed that KIF26B antisense RNA 1 (KIF26B-AS1) propels cell proliferation and migration in laryngeal cancer and regulates the toll-like receptor 4 (TLR4) signaling pathway. KIF26B-AS1 also recruits FUS to stabilize TLR4 mRNA, consequently activating the TLR4 signaling pathway. Furthermore, KIF26B-AS1 plays an oncogenic role in laryngeal cancer via upregulating TLR4 expression as well as the FUS/TLR4 pathway axis, findings which offer novel insight for targeted therapies in the treatment of laryngeal cancer patients.

Synergistic effect of yeast integrated with alkyl polyglucose for short-chain fatty acids production from sludge anaerobic fermentation.

An efficient strategy for short-chain fatty acid (SCFA) production from sludge anaerobic fermentation was proposed with the combination of yeast and alkyl polyglucose (APG). It revealed that the synergetic effect of yeast and APG could boost the SCFA concentration to the maximum value of 2800.34 mg COD/L within 9 days at 0.20 g/g suspended solids (SS) yeast and 0.20 g/g SS APG, which was significantly higher than that of its counterparts. Interestingly, the sludge solubilization, the biodegradability of fermentation substrate, as well as the acidification of hydrolyzed products, was evidently improved in the coexistence of APG and yeast. The activities of hydrolytic enzymes and acetate kinase were also stimulated, whereas the coenzyme F420 was inhibited. The synergetic effect of yeast and APG used in this work enriches the study of carbon resource recovery from sludge anaerobic fermentation.

Aging of polyvinylidene fluoride (PVDF) ultrafiltration membrane due to ozone exposure in water treatment: Evolution of membrane properties and performance.

Pre-ozonation is an effective pretreatment tactic for mitigating fouling of ultrafiltration (UF) membrane in water and wastewater treatment, but the compatibility of polymeric UF membranes with residual ozone remains unclear. In this study, effects of long-term ozone exposure on properties and performance of polyvinylidene fluoride (PVDF) UF membrane reinforced by polyethylene terephthalate (PET) layer were systematically investigated. The exposure intensities were designed to simulate ozone exposure at 0.1 mg/L for 0.5-5 years. Chemical composition analysis suggested that the hydrophilic additives, such as possibly polyvinyl pyrrolidone (PVP), was gradually degraded and released from the membrane, whereas the PVDF matrix exhibited fairly good ozone resistance. Ozonation resulted in increase of pore size and decrease of surface hydrophilicity, which can be attributed to oxidation and dislodgement of hydrophilic additives. Accordingly, long-term ozonation led to moderate changes in performance factors, including increase of membrane permeability by 34%, decrease of retention ability by 21.8%, increase of organic fouling propensity. It is worth noting that membrane tensile strength suffered substantial decrease after ozonation, probably due to ozonation of the PET support layer. Overall, it seems that the PVDF functional layer exhibited good ozone resistance, but the PET support layer was the Achilles' heel of the reinforced PVDF membrane for integrating with pre-ozonation.

Effects of reward associations on components of attentional network.

Previous research has revealed the influence of reward associations on attentional selection and control. The attentional network can be divided into three components according to its function: Alerting, orienting, and executive control. In the current research, we used training-test procedure and attention network test variant to investigate the effects of color-based reward associations on alerting (Experiment 1), orienting (Experiment 2), executive control (Experiment 3), as well as the interactions among these three networks (Experiment 4). The findings were as follows: Compared with colors previously associated with low reward, colors previously associated with high reward trigger stronger alerting and orienting effects (Experiments 1 and 2), and they had stronger interference effects when functioning as features of flanker distractor (Experiment 3). More importantly, reward associations had only a positive impact on the interaction of orienting by executive control but not on the interaction of alerting by executive control (Experiment 4). In summary, reward associations have different effects on the three attentional networks and can enhance the interaction of orienting by executive control.

[Influence of Secondary Organic Coating on Hygroscopicity of a Sodium Chloride Core: Based on Mircro-scale Single Particle Analysis].

The hygroscopicity of atmospheric particulate matter can determine its optical behaviors, cloud condensation nuclei activity, and heterogeneous chemical reaction on the surface. However, the effect of secondary organic matter on the hygroscopicity of inorganic salts is still rarely studied. In this study, a 2 L gas-phase flow tube was used to stimulate the oxidation process of α-pinene. NaCl particles at the end of the flow tube served as seeds to absorb the product of α-pinene oxidation. The absorption times were 11, 15, and 20 h. Transmission electron microscopy results show that this system could produce particles with typical core-shell structure, and that the thickness of the organic shell would be thicker with longer collection time. Results from individual particle hygroscopic systems indicate that the organic shell can affect the deliquescent point of NaCl (pure NaCl deliquesces at 77%):for 11 h collection time, the NaCl core began to deliquesce at 75.5% relative humidity, and completely deliquesced at 78%; for 15 h collection time, the deliquescent range of the NaCl core was 75%-78.5%; and the deliquescent range was 76%-83% for the NaCl core with 20 h collection time. Furthermore, the organic shell could affect the hygroscopic growth of the NaCl core:the growth factor of the NaCl core with 11 h, 15 h, and 20 h collection time was 2.5, 2.3, and 1.8, respectively. These results indicate that:① the secondary organic shell could cause water absorption of the NaCl core in advance:② the organic shell could postpone the completely deliquescent point of the NaCl core, and this effect is more obvious with a thicker organic shell; ③ the organic shell could restrain the hygroscopic growth factor of the NaCl core. This study suggests that it is necessary to consider the influence of secondary organic matter on inorganic salts when studying the hygroscopic properties of atmospheric particles in climate models and field observations.

The characterization of lung microbiome in lung cancer patients with different clinicopathology.

There were few knowledge concerned correlation between lung microbiome and different clinicopathology of lung cancer. Bronchial washing fluid (BWF) and sputum are commonly used sample types but there was no study comparing difference of microbiome between these two in lung cancer. In this study, we aimed to compare difference of microbiome between these two sample types and characterize lung microbiome in squamous cell lung carcinoma with (SCC_M1) or without distant metastasis (SCC_M0) and lung adenocarcinoma with (AD_M1) or without distant metastasis (AD_M0). We collected 40 BWF samples and 52 sputum samples from newly diagnosed lung cancer patients. Bacterial species were sequenced via 16S rRNA sequencing. Phylum Proteobacteria in BWF samples were significantly higher than sputum samples (Wilcoxon test, P = 0.003). At phylum level, microbiome of BWF samples was more similar to that of lung cancer tissues reported in the previous literature. LEFse analysis showed that in BWF group, genera Veillonell, Megasphaera, Actinomyces and Arthrobacter in AD_M0 were significantly higher than those in SCC_M0, and genera Capnocytophaga and Rothia in AD_M1 were significantly lower than that in SCC_M1. Compared with AD_M0, genus Streptococcus of AD_M1 was significantly lower, and genera Veillonella and Rothia in SCC_M1 were significantly higher than that in SCC_M1. Our study suggested that BWF samples might better reflect the microbiome of lung cancer tissues. In different metastatic states of lung cancer, differential genera between squamous cell carcinoma and adenocarcinoma were different. And in different histologic types of lung cancer, distant metastasis-related genera were not the same.

Chemical Cleaning of Ultrafiltration Membrane Fouled by Humic Substances: Comparison between Hydrogen Peroxide and Sodium Hypochlorite.

Chemical cleaning is indispensable for the sustainable operation of ultrafiltration (UF) system in water and wastewater treatment. Sodium hypochlorite (NaClO) is an established cleaning agent for membranes subject to organic and microbial fouling, but concerns have been raised about the generation of toxic halogenated by-products during NaClO cleaning. Hydrogen peroxide (H2O2) is a potential "green" cleaning agent that can avoid the formation of halogenated by-products. In this work, cleaning efficacy of H2O2 and NaClO for UF membrane fouled by humic substances (HS) was evaluated under a wide pH range, and change of HS's properties due to reaction with cleaning agents was examined. The cleaning efficacy of H2O2 was lower than that of NaClO at pH 3-9, but it increased to a level (91.4%) comparable with that of NaClO at pH 11. The extents of changes in properties and fouling potential of HS due to reacting with cleaning agents were consistent with their cleaning efficacy. H2O2 treatment at pH 11 significantly increased negative charge of HS molecules, decomposed high-MW molecules, and reduced its fouling potential. Therefore, considering treatment/disposal of cleaning waste and cleaning efficacy, H2O2 cleaning under strong alkaline condition can be a good choice for HS-fouled membrane.

Hydrodynamic shear force shaped the microbial community and function in the aerobic granular sequencing batch reactors for low carbon to nitrogen (C/N) municipal wastewater treatment.

The lab-scale aerobic granules process was applied for low carbon to nitrogen (C/N < 4) wastewater treatment under different hydrodynamic shear forces. Results revealed that aerobic granules exhibited strong adaptability and stability. The aerobic granules might adopt an extracellular polymeric substances (EPS) regulating mechanism to address the changes in operational conditions, especially through growing secretion of fluorescence protein. The hydrodynamic shear force determinedly shaped and regulated the diversity and structure of dominant microbial community, briefly, reduced aeration intensity with increased time led to higher microbial richness, lower diversity and evenness, and shifts of predominant microorganisms. Phylogenetic classification of the key functional groups including bacteria related to carbon and nutrients removal, EPS production and quorum sensing (QS) presented much more differences among the reactors subject to different conditions. Therefore, the present work adds insight into the comprehensive understanding of the effect of aeration induced hydrodynamic shear force on aerobic granules.

Natural sunlight induced rapid formation of water-born algal-bacterial granules in an aerobic bacterial granular photo-sequencing batch reactor.

Wastewater treatment by means of algal-bacterial granules has become a hot topic worldwide recently. Rapid granulation of algal-bacterial granules was achieved in an aerobic bacterial granular sequencing batch reactor (SBR) under natural sunlight exposure. Occurrence of abundant filamentous bacteria bridging the water-born algae, and overproduction of extracellular polymeric substances (EPS) (especially polysaccharides (PS), tryptophan & protein-like, and humic acid-like substances) were observed on the first 3 days, while the algae grew into the inner side of the granules and mature granules were obtained on day 7. The growth of the water-born algae slightly decreased the settleability, mean sizes of the granules, but stimulated the bioactivity significantly. Whereas, the biomass retention decreased before day 3, and got stable soon with the maturation period with symbiotic growth of algal-bacterial biomass. Illumina results revealed that the introduction of algae reduced the richness and diversity of bacterial community. Besides, few changes in structure and some compositions shifts in bacterial communities were found, while the predominant algae shifted from Diatomea to green algae Chlorophyceae. The possible mechanism for natural sunlight induced granulation of algal-bacterial granules was thus proposed based on the interactions between algae and bacteria.

Comparison of the Efficacy, Side Effects, and Cost of Modafinil and Intranasal Mometasone Furoate in Obstructive Sleep Apnea-Hypopnea Syndrome: A Preliminary Clinical Study.

BACKGROUND Obstructive sleep apnea-hypopnea syndrome (OSAHS) is characterized by repeated episodes of reduction in airflow due to the collapse of the upper airway during sleep. The aim of this study was to compare clinical outcome, side effects, and cost of treatment between modafinil and intranasal mometasone furoate in patients with OSAHS. MATERIAL AND METHODS Patients with OSAHS (N=250) were divided into two groups: the modafinil group (MG) (N=125) were treated with 100 mg modafinil twice a day; the intranasal mometasone furoate group (IMFG) (N=125) were treated with 100 µg of intranasal mometasone furoate in the evening. Quality of life, grading of OSAHS, plain-film radiography, the adenoidal-nasopharyngeal ratio (AN ratio), side effects, cost of treatment, and beneficial effects after discontinuation of treatment were evaluated for all patients. RESULTS Duration of sleep apnea was significantly reduced in the IMFG compared with the MG (p=0.0145, q=9.262). Modafinil and intranasal mometasone furoate both had moderate effects on improvement of the OSAHS score. The IMFG showed a significantly greater beneficial effect on the AN ratio when compared with the MG (p=0.0001, q=6.584). No adverse events of treatment with modafinil and intranasal mometasone furoate were reported. Cost of treatment and beneficial effect after discontinuation were both significantly greater for the IMFG compared with the MG. CONCLUSIONS The findings of this preliminary clinical study were that for patients diagnosed with OSAHS, night-time treatment with intranasal mometasone furoate was more effective than modafinil.

Simultaneous nitrification, denitrification and phosphorus removal in aerobic granular sequencing batch reactors with high aeration intensity: Impact of aeration time.

A new operating approach by reducing the aeration time while keeping high intensity was evaluated for enhanced nutrients removal and maintenance of granular stability. Three aerobic granular sequencing batch reactors (SBR) performing simultaneous nitrification, denitrification and phosphorus removal (SNDPR) were run at different aeration time (120, 90, and 60 min). Aerobic granules could remain their integrity and stability over long-term operation under high aeration intensity and different time, and shorter aeration time favored the retention of biomass, better settleability, and more production of extracellular polymeric substances (EPS). Besides, efficient and stable reactor performance for carbon and phosphorus were achieved, especially, enhanced nitrogen removal was obtained due to reduction of aeration time. Further exploration revealed that the aeration time shaped the bacterial community in terms of diversity, composition, as well as the distribution of functional groups involving carbon, nitrogen and phosphorus removal.

Macrophage responses to the physical burden of cell-sized particles.

The role of a biophysical signal on cell response has excited tremendous interest recently. Herein, we exploited an "intake method" together with uniform autofluorescent cell-sized particles to investigate macrophage responses against different particle burdens. Our work not only revealed an insatiable macrophage uptake of cell-sized microparticles (MPs) but also widened the theoretical size and volume range for particle entry. MPs outperform NPs in the utmost volume of intracellular particles, indicating a converse size-associated event than originally anticipated. Such a superior volume burden (2.64-fold of the cell volume) for MPs is highly correlated to less membrane loss (19-fold below that for NPs) and more evident deformation of the cell membrane/nucleus. In addition, the cells with a high burden of MPs exhibit moderate migration activity, which is in line with mild cytokine release. These results highlight the indispensable role of physical burden on the regulation of macrophage functions, providing important views for desired biological outcomes and minimal side effects.

Systematic CRISPR-Cas9-Mediated Modifications of Plasmodium yoelii ApiAP2 Genes Reveal Functional Insights into Parasite Development.

Malaria parasites have a complex life cycle with multiple developmental stages in mosquito and vertebrate hosts, and different developmental stages express unique sets of genes. Unexpectedly, many transcription factors (TFs) commonly found in eukaryotic organisms are absent in malaria parasites; instead, a family of genes encoding proteins similar to the plant Apetala2 (ApiAP2) transcription factors is expanded in the parasites. Several malaria ApiAP2 genes have been shown to play a critical role in parasite development; however, the functions of the majority of the ApiAP2 genes remain to be elucidated. In particular, no study on the Plasmodium yoelii ApiAP2 (PyApiAP2) gene family has been reported so far. This study systematically investigated the functional roles of PyApiAP2 genes in parasite development. Twenty-four of the 26 PyApiAP2 genes were selected for disruption, and 12 were successfully knocked out using the clustered regularly interspaced short palindromic repeat-CRISPR-associated protein 9 (CRISPR-Cas9) method. The effects of gene knockout (KO) on parasite development in mouse and mosquito stages were evaluated. Ten of 12 successfully disrupted genes, including two genes that have not been functionally characterized in any Plasmodium species previously, were shown to be critical for P. yoelii development of sexual and mosquito stages. Additionally, seven of the genes were labeled for protein expression analysis, revealing important information supporting their functions. This study represents the first systematic functional characterization of the P. yoelii ApiAP2 gene family and discovers important insights on the roles of the ApiAP2 genes in parasite development.IMPORTANCE Malaria is a parasitic disease that infects hundreds of millions of people, leading to an estimated 0.35 million deaths in 2015. A better understanding of the mechanism of gene expression regulation during parasite development may provide important clues for disease control and prevention. In this study, systematic gene disruption experiments were performed to study the functional roles of members of the Plasmodium yoelii ApiAP2 (PyApiAP2) gene family in parasite development. Genes that are critical for the development of male and female gametocytes, oocysts, and sporozoites were characterized. The protein expression profiles for seven of the PyApiAP2 gene products were also analyzed, revealing important information on their functions. This study provides expression and functional information for many PyApiAP2 genes, which can be explored for disease management.

[Diffusion of fluorescent and magnetic molecular probes in brain interstitial space].

OBJECTIVE: To compare the diffusion properties of fluorescent probes dextran-tetramethylrhodamine (DT) and lucifer yellow CH (LY) and magnetic probe gadolinium-diethylene triamine pentaacetic acid (Gd-DTPA) in porous media and to screen out a suitable fluorescent probe for optical imaging of brain interstitial space (ISS). METHODS: Agarose gels sample were divided into DT group, LY group and Gd-DTPA group, and the corresponding molecular probes were imported in each group. The dynamic diffusions of DT and LY in agarose gels at different time points (15, 30, 45, 60, 90, and 120 min) were scanned with laser scanning confocal microscope, the dynamic diffusion of Gd-DTPA was imaged with magnetic resonance imaging. The average diffusion speed of LY were demonstrated to be consistent with those of Gd-DTPA. The LY was introduced into caudate putamen of 18 rats, respectively, the diffusion of LY in the sequential slices of rat brain at different time points (0.5, 1, 2, 3, 7, 11 h) were scanned, and the results were compared with those of rats' brain with Gd-DTPA imported and imaged in vivo with magnetic resonance imaging. RESULTS: The diffusions of the three probes were isotropic in the agarose gels, and the average diffusion speeds of DT, LY and Gd-DTPA were: (0.07±0.02)×10(-2) mm2/s, (1.54±0.47)×10(-2) mm2/s, (1.45±0.50)×10(-2) mm2/s, respectively. The speed of DT was more slower than both LY and Gd-DTPA (ANOVA, F=367.15, P<0.001; Post-Hoc LSD, P<0.001), and there was no significant difference between the speeds of LY and Gd-DTPA (Post-Hoc LSD, P=0.091). The variation tendency of diffusion area of DT was different with both that of LY and that of Gd-DTPA (Bonferroni correction, α=0.0125, P<0.001), and there was no significant difference between LY and Gd-DTPA (Bonferroni correction, α=0.0125, P=0.203), in analysis by repeated measures data of ANOVA. The diffusions of LY and Gd-DTPA were anisotropy in rat caudate putamen,and the average diffusion speeds of LY and Gd-DTPA were: (1.03±0.29)×10(-3) mm2/s, (0.81±0.27)×10(-3) mm2/s, respectively, no significant difference was demonstrated (t=0.759, P=0.490); half-time of single intensity of LY and Gd-DTPA was (2.58±0.04) h, (2.46±0.10) h, respectively, no significant difference was found (t=2.025, P=0.113). The diffusion area ratios between LY and Gd-DTPA in rat caudate putamen was not statistically different at hours 0.5, 1, 2, 3 and 7 (t=2.249, P=0.088; t=2.582, P=0.061; t=1.966, P=0.121; t=0.132, P=0.674; t=0.032, P=0.976), while, a slightly difference was found at 11 h (t=2.917, P=0.043,in analysis by t test). CONCLUSION: LY present the same diffusion property with Gd-DTPA in porous media witch including agarose gels and live rat brain tissue, indicates that LY is a suitable fluorescent probe for optical imaging of brain ISS, and it can be used for microscopic, macro and in vitro measure of brain ISS.

[The correlation analysis of coagulation detection and blood routine parameters of sudden hearing loss].

OBJECTIVE: Through the analysis of coagulation convention and blood routine parameters of sudden hearing loss (SHL) patients, further prove the correlation of sudden deafness and the the inner ear microcirculation, to guide clinical diagnosis and treatment. METHOD: Select 424 patients (448 ears) with sudden deafness in our department to SHL group. According to hearing curve is classified into low intermediate frequency descent group, high frequency drop and full frequency group, and drawing 244 cases in the same period of hospitalization deviated septum, vocal cord polyp patients as control group. All patients' coagulation detection, D-dimer, blood leukocytes, neutrophils and platelet count percentages were analyzed. Then a meaningful factor multivariate Logistic regression analysis was made. RESULT: There was a statistically significant difference between the two groups' prothrombin time, international normalized ratio, activated partial thromboplastin time, thrombin time measurement, fibrinogen, D-dimer, platelet count, white blood cell, neutrophil ratio(P<0.05); Logistic regression analysis showed that the prothrombin, thrombin time measurement, fibrinogen, D-dimer, neutrophil incidence of sudden hearing loss associated risk factors. CONCLUSION: SHL in patients with coagulation dysfunction may be involved in the occurrence of SHL development mechanism, and there is a correlation of the SHL and the dysfunction of inner ear microcirculation.

Bioinformatics analysis of proteomic profiles during the process of anti-Thy1 nephritis.

Anti-Thy1 nephritis is a well-established experimental mesangial proliferative nephritis model. Exploring the molecular mechanisms of pathophysiology in anti-Thy1 nephritis may elucidate the pathogeneses of mesangial proliferation. We examined the roles and acting mechanisms of differentially expressed proteins (DEPs) by bioinformatics analysis of glomeruli proteomic profiles during the course of anti-Thy1 nephritis. In total, 108 DEPs were found by two-dimensional fluorescence difference gel electrophoresis (2D-DIGE), and 40 DEPs were identified by matrix-assisted laser desorption ionization/time of flight and liquid chromatography-MS. DEPs were classified into five clusters (Clusters 1-5), according to their expression trends using Cluster 3.0 software, involved in regulating biological processes such as the stress response, cell proliferation, apoptosis, energy metabolism, transport, and the actin cytoskeleton. The expression patterns of ten DEPs, distributed across five clusters, including AKR1A1, AGAT, ATP6V1B2, HIBADH, MDH1, MPST, NIT2, PRDX6, PSMB7, and TPI1, were validated by Western blotting. Based on Western blotting and immunohistochemistry, we also found that the DEP FHL2, which was primarily expressed in the mesangial region, was down-regulated on days 3 and 5, and up-regulated on day 10. In vitro, we found that FHL2 overexpression induced mesangial cell proliferation by increasing the number of S-phase cells and decreasing G2/M-phase cells, whereas inhibiting FHL2 had the opposite effect. This study explored novel DEPs and their expression patterns during anti-Thy1 nephritis, and elucidated FHL2's effect on mesangial cell proliferation. These results will contribute to our understanding of the pathogenesis of mesangial proliferation.

Change of MAX interactor 1 expression in an anti-Thy1 nephritis model and its effect on mesangial cell proliferation.

BACKGROUND/AIMS: During the disease process of mesangial proliferative glomerulonephritis, the expression of various factors that influence mesangial proliferation is altered. MAX interactor 1 (Mxi1) antagonizes the transcription factor Myc and is believed to be a tumor suppressor. However, no studies have investigated its effect on mesangial cell proliferation. METHODS: To investigate the effect of Mxi1 on renal mesangial cell proliferation, we established a classic rat anti-Thy1 mesangial proliferative glomerulonephritis model. Mesangial proliferation was estimated by immunohistochemical analysis of Ki67. Mxi1 expression at each time point was assessed by real-time RT-PCR and Western blot analyses. Furthermore, we altered the expression level of Mxi1 by a plasmid and siRNA to detect its effect on rat mesangial cell proliferation in vitro. RESULTS: Mxi1 expression decreased significantly during the proliferative period of anti-Thy1 nephritis model and then gradually increased as proliferation declined, indicating that Mxi1 may be linked to mesangial cell proliferation. Upregulation of Mxi1 expression via plasmid transfection in vitro reduced the expression of the positive-acting cell cycle regulatory proteins cyclin B1, cyclin D1, cyclin E, CDC2 and CDK2; significantly reduced mesangial cell proliferation; reduced the percentage of S phase cells; and increased the percentage of G2/M phase cells. Inhibition of Mxi1 expression by siRNA in vitro produced the opposite effects: increased expression of cyclin B1, cyclin D1, cyclin E, CDC2 and CDK2; markedly increased cell proliferation; higher percentage of S phase cells; and dramatically lower percentage of G2/M phase cells. Transcription factor c-myc protein expression showed no obvious difference after Mxi1 plasmid and siRNA transfection. The expressions of cell cycle regulatory proteins mentioned above were negative correlated with Mxi1 expression in anti-Thy1 nephritis model. CONCLUSION: These results suggest that Mxi1 expression levels were inversely correlated with proliferation in anti-Thy1 nephritis rats and it may influence cell cycle progression and thus the rate of mesangial cell proliferation by regulating the expression of c-myc target cell cycle regulatory proteins.