Integrated transcriptomic and metabolomic analyses reveals anthocyanin biosynthesis in leaf coloration of quinoa (Chenopodium quinoa Willd.).

BACKGROUND: Quinoa leaves demonstrate a diverse array of colors, offering a potential enhancement to landscape aesthetics and the development of leisure-oriented sightseeing agriculture in semi-arid regions. This study utilized integrated transcriptomic and metabolomic analyses to investigate the mechanisms underlying anthocyanin synthesis in both emerald green and pink quinoa leaves. RESULTS: Integrated transcriptomic and metabolomic analyses indicated that both flavonoid biosynthesis pathway (ko00941) and anthocyanin biosynthesis pathway (ko00942) were significantly associated with anthocyanin biosynthesis. Differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) were analyzed between the two germplasms during different developmental periods. Ten DEGs were verified using qRT-PCR, and the results were consistent with those of the transcriptomic sequencing. The elevated expression of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), 4-coumarate CoA ligase (4CL) and Hydroxycinnamoyltransferase (HCT), as well as the reduced expression of flavanone 3-hydroxylase (F3H) and Flavonol synthase (FLS), likely cause pink leaf formation. In addition, bHLH14, WRKY46, and TGA indirectly affected the activities of CHS and 4CL, collectively regulating the levels of cyanidin 3-O-(3'', 6''-O-dimalonyl) glucoside and naringenin. The diminished expression of PAL, 4CL, and HCT decreased the formation of cyanidin-3-O-(6"-O-malonyl-2"-O-glucuronyl) glucoside, leading to the emergence of emerald green leaves. Moreover, the lowered expression of TGA and WRKY46 indirectly regulated 4CL activity, serving as another important factor in maintaining the emerald green hue in leaves N1, N2, and N3. CONCLUSION: These findings establish a foundation for elucidating the molecular regulatory mechanisms governing anthocyanin biosynthesis in quinoa leaves, and also provide some theoretical basis for the development of leisure and sightseeing agriculture.

Enhancement of broad-spectrum disease resistance in wheat through key genes involved in systemic acquired resistance.

Systemic acquired resistance (SAR) is an inducible disease resistance phenomenon in plant species, providing plants with broad-spectrum resistance to secondary pathogen infections beyond the initial infection site. In Arabidopsis, SAR can be triggered by direct pathogen infection or treatment with the phytohormone salicylic acid (SA), as well as its analogues 2,6-dichloroisonicotinic acid (INA) and benzothiadiazole (BTH). The SA receptor non-expressor of pathogenesis-related protein gene 1 (NPR1) protein serves as a key regulator in controlling SAR signaling transduction. Similarly, in common wheat (Triticum aestivum), pathogen infection or treatment with the SA analogue BTH can induce broad-spectrum resistance to powdery mildew, leaf rust, Fusarium head blight, and other diseases. However, unlike SAR in the model plant Arabidopsis or rice, SAR-like responses in wheat exhibit unique features and regulatory pathways. The acquired resistance (AR) induced by the model pathogen Pseudomonas syringae pv. tomato strain DC3000 is regulated by NPR1, but its effects are limited to the adjacent region of the same leaf and not systemic. On the other hand, the systemic immunity (SI) triggered by Xanthomonas translucens pv. cerealis (Xtc) or Pseudomonas syringae pv. japonica (Psj) is not controlled by NPR1 or SA, but rather closely associated with jasmonate (JA), abscisic acid (ABA), and several transcription factors. Furthermore, the BTH-induced resistance (BIR) partially depends on NPR1 activation, leading to a broader and stronger plant defense response. This paper provides a systematic review of the research progress on SAR in wheat, emphasizes the key regulatory role of NPR1 in wheat SAR, and summarizes the potential of pathogenesis-related protein (PR) genes in genetically modifying wheat to enhance broad-spectrum disease resistance. This review lays an important foundation for further analyzing the molecular mechanism of SAR and genetically improving broad-spectrum disease resistance in wheat.

AsHSP26.2, a creeping bentgrass chloroplast small heat shock protein positively regulates plant development.

KEY MESSAGE: The creeping bentgrass small heat shock protein AsHSP26.2 positively regulates plant growth and is a novel candidate for use in crop genetic engineering for enhanced biomass production and grain yield. Small heat shock proteins (sHSPs), a family of proteins with high level of diversity, significantly influence plant stress tolerance and plant development. We have cloned a creeping bentgrass chloroplast-localized sHSP gene, AsHSP26.2 responsive to IAA, GA and 6-BA stimulation. Transgenic creeping bentgrass overexpressing AsHSP26.2 exhibited significantly enhanced plant growth with increased stolon number and length as well as enlarged leaf blade width and leaf sheath diameters, but inhibited leaf trichomes initiation and development in the abaxial epidermis. These phenotypes are completely opposite to those displayed in the transgenic plants overexpressing AsHSP26.8, another chloroplast sHSP26 isoform that contains additional seven amino acids (AEGQGDG) between the consensus regions III and IV (Sun et al., Plant Cell Environ 44:1769-1787, 2021). Furthermore, AsHSP26.2 overexpression altered phytohormone biosynthesis and signaling transduction, resulting in elevated auxin and gibberellins (GA) accumulation. The results obtained provide novel insights implicating the sHSPs in plant growth and development regulation, and strongly suggest AsHSP26.2 to be a novel candidate for use in crop genetic engineering for enhanced plant biomass production and grain yield.

The impact of digital platforms on new startup performance: Strategy as moderator.

Recent studies have acknowledged the contribution of digital platforms to entrepreneurial activity. However, little has been discovered regarding the way digital platforms impact new start-ups' performance and how strategy affects this relationship. This study investigated this phenomenon using a structural equation model based on entrepreneurial bricolage and environmental strategy performance theory. The current findings indicate that broader market outreach, cost-effectiveness, and network effect produce a favorable, direct, and substantial effect on new startup performance. Furthermore, strategy moderates the effects of broader market outreach, network effect, and cost-effectiveness on new startup performance. Furthermore, strategy moderates the impact of broader market outreach, network effect, and cost-effectiveness on new startup performance. The Chinese government in particular, and governments of various emerging nations generally, must pay close attention to the results of this research to design policies that will not only lengthen the lifespan of small businesses but also promote rapid expansion as well as innovation of digital start-ups.

Biological interactions of polystyrene nanoplastics: Their cytotoxic and immunotoxic effects on the hepatic and enteric systems.

As emerging pollutants in the environment, nanoplastics (NPs) can cross biological barriers and be enriched in organisms, posing a greatest threat to the health of livestock and humans. However, the size-dependent toxic effects of NPs in higher mammals remain largely unknown. To determine the size-dependent potential toxicities of NPs, we exposed mouse (AML-12) and human (L02) liver cell lines in vitro, and 6-week-old C57BL/6 mice (well-known preclinical model) in vivo to five different sizes of polystyrene NPs (PS-NPs) (20, 50, 100, 200 and 500 nm). We found that ultra-small NPs (20 nm) induced the highest cytotoxicity in mouse and human liver cell lines, causing oxidative stress and mitochondrial membrane potential loss on AML-12 cells. Unexpectedly in vivo, after long-term oral exposure to PS-NPs (75 mg/kg), medium NPs (200 nm) and large NPs (500 nm) induced significant hepatotoxicity, evidenced by increased oxidative stress, liver dysfunction, and lipid metabolism disorders. Most importantly, medium or large NPs generated local immunotoxic effects via recruiting and activating more numbers of neutrophils and monocytes in the liver or intestine, which potentially resulted in increased proinflammatory cytokine secretion and the tissue damage. The discrepancy in in vitro-in vivo toxic results might be attributed to the different properties of biodistribution and tissue accumulation of different sized NPs in vivo. Our study provides new insights regarding the hepatotoxicity and immunotoxicity of NPs on human and livestock health, warranting us to take immense measures to prevent these NPs-associated health damage.

Diversity in boardroom and debt financing: A case from China.

The study aims to explore the role of gender diversity in debt financing choices among Chinese listed firms. The study used the Chinese listed firm's data from 1991 to 2022 from the Chinese Stock Market return. The study used the fixed effect regression analysis and revealed that gender diversity positively affects debt financing among Chinese firms. Additionally, mass theory results suggested that at least three females on the board significantly influence firms. It served as the voice of gender diversity to influence the board's decisions regarding debt financing. The study has several theoretical and practical implications. This study will enlighten the Chinese boardroom dynamics by reassuring them to add more females to diversity policies. It will benefit future studies on boardroom activities and debt financing in emerging economies. It will be practical guidance for the Chinese policymakers, governing authorities, and corporate executives. The study stresses the need for significant diversity on the board rather than one female presence on the board. Secondly, this study contradicts the stereotype perception that females are not making risky decisions.

Two novel truncating variants in UBAP1 are responsible for hereditary spastic paraplegia.

Hereditary spastic paraplegias (HSPs) are a group of rare neurodegenerative disorders. HSPs are complex disorders and are clinically and genetically heterogeneous. To date, more than 80 genes or genetic loci have been reported to be responsible for HSPs in a Mendelian-dependent manner. Most recently, ubiquitin-associated protein 1 (UBAP1) has been recognized to be involved in HSP. Here, we identified novel protein truncating variants in two families with pure form of HSP. A novel deletion (c.468_469delTG) in the UBAP1 gene was found in the first family, whereas a nonsense variant (c.512T>G) was ascertained in the second family. The variants were confirmed in all patients but were not detected in unaffected family members. The mutations resulted in truncated proteins of UBAP1. The variants did not result in different subcellular localizations in neuro-2a cells. However, each of the two variants impaired neurite outgrowth. Taken together, our findings expand the pathogenic spectrum of UBAP1 variants in HSP.

MicroRNA-144 Suppresses Prostate Cancer Growth and Metastasis by Targeting EZH2.

BACKGROUND: Prostate cancer is a common malignant tumor with a high incidence. MicroRNAs (miRNAs) have been shown to be important post-transcriptional regulators during tumorigenesis. This study aimed to explore the effect of miR-144 on PCa proliferation and apoptosis. MATERIAL AND METHODS: The expression of miR-144 and EZH2 were examined in clinical PCa tissues. PCa cell line LNCAP and DU-145 was employed and transfected with miR-144 mimics or inhibitors. The correlation between miR-144 and EZH2 was verified by luciferase reporter assay. Cell viability, apoptosis and migratory capacity were detected by CCK-8, flow cytometry assay and wound healing assay. The protein level of EZH2, E-Cadherin, N-Cadherin and vimentin were analyzed by western blotting. RESULTS: miR-144 was found to be negatively correlated to the expression of EZH2 in PCa tissues. Further studies identified EZH2 as a direct target of miR-144. Moreover, overexpression of miR-144 downregulated expression of EZH2, reduced cell viability and promoted cell apoptosis, while knockdown of miR-144 led to an inverse result. miR-144 also suppressed epithelial-mesenchymal transition level of PCa cells. CONCLUSION: Our study indicated that miR-144 negatively regulate the expression of EZH2 in clinical specimens and in vitro. miR-144 can inhibit cell proliferation and induce cell apoptosis in PCa cells. Therefore, miR-144 has the potential to be used as a biomarker for predicting the progression of PCa.

A chloroplast heat shock protein modulates growth and abiotic stress response in creeping bentgrass.

Small heat shock proteins (sHSPs), a family of the ubiquitous stress proteins in plants acting as molecular chaperones to protect other proteins from stress-induced damage, have been implicated in plant growth and development as well as plant response to environmental stress, especially heat stress. In this study, a chloroplast-localized sHSP, AsHSP26.8, was overexpressed in creeping bentgrass (Agrostis stolonifera L.) to study its role in regulating plant growth and stress response. Transgenic (TG) creeping bentgrass plants displayed arrested root development, slow growth rate, twisted leaf blades and are more susceptible to heat and salt but less sensitive to drought stress compared to wild-type (WT) controls. RNA-seq analysis revealed that AsHSP26.8 modulated the expression of genes in auxin signalling and stress-related genes such as those encoding HSPs, heat shock factors and other transcription factors. Our results provide new evidence demonstrating that AsHSP26.8 negatively regulates plant growth and development and plays differential roles in plant response to a plethora of diverse abiotic stresses.

Therapeutic Validation of Venous Pulsatile Tinnitus and Biomaterial Applications for Temporal Bone Reconstruction Surgery Using Multi-sensing Platforms and Coupled Computational Techniques.

The application of grafts and biomaterials is a cardinal therapeutic procedure to resolve venous pulsatile tinnitus (PT) caused by temporal bone dehiscence during transtemporal reconstructive surgery. However, the transmission mechanism of venous PT remains unclear, and the sound absorption and insulation properties of different repair materials have not been specified. This study quantifies the vibroacoustic characteristics of PT, sources the major transmission pathway of PT, and verifies the therapeutic effect of different material applications using joint multi-sensing platforms and coupled computational fluid dynamics (CFD) techniques. The in vivo intraoperative acoustic and vibroacoustic characteristics of intrasinus blood flow motion and dehiscent sigmoid plate of a typical venous PT patient were investigated using acoustic and displacement sensors. The acoustical, morphological, and mechanical properties of the dehiscent sigmoid plate, grafts harvested from a cadaveric head, and other biomaterials were acquired using acoustical impedance tubes, micro-CT, scanning electron microscopy, and mercury porosimetry, as appropriate. To analyze the therapeutic effect of our previous reconstructive techniques, coupled CFD simulations were performed using the acquired mechanical properties of biomaterials and patient-specific radiologic data. The peak in vivo intraoperatively gauged, peak simulated vibroacoustic and peak simulated hydroacoustic amplitude of PT prior to sigmoid plate reconstruction were 64.0, 70.4, and 72.8 dB, respectively. After the solidified gelatin sponge-bone wax repair technique, the intraoperative gauged peak amplitude of PT was reduced from 64.0 to 47.3 dB. Among three different reconstructive techniques based on CFD results, the vibroacoustic and hydroacoustic sounds were reduced to 65.9 and 68.6 dB (temporalis-cartilage technique), 63.5 and 63.1 dB (solidified gelatin sponge technique), and 42.4 and 39.2 dB (solidified gelatin sponge-bone wax technique). In conclusion, the current novel biosensing applications and coupled CFD techniques indicate that the sensation of PT correlates with the motion and impact from venous flow, causing vibroacoustic and hydroacoustic sources that transmit via the air-conduction transmission pathway. The transtemporal reconstructive surgical efficacy depends on the established areal density of applied grafts and/or biomaterials, in which the total transmission loss of PT should surpass the amplitude of the measured loudness of PT.

AsHSP26.8a, a creeping bentgrass small heat shock protein integrates different signaling pathways to modulate plant abiotic stress response.

BACKGROUND: Small heat shock proteins (sHSPs) are critical for plant response to biotic and abiotic stresses, especially heat stress. They have also been implicated in various aspects of plant development. However, the acting mechanisms of the sHSPs in plants, especially in perennial grass species, remain largely elusive. RESULTS: In this study, AsHSP26.8a, a novel chloroplast-localized sHSP gene from creeping bentgrass (Agrostis stolonifera L.) was cloned and its role in plant response to environmental stress was studied. AsHSP26.8a encodes a protein of 26.8 kDa. Its expression was strongly induced in both leaf and root tissues by heat stress. Transgenic Arabidopsis plants overexpressing AsHSP26.8a displayed reduced tolerance to heat stress. Furthermore, overexpression of AsHSP26.8a resulted in hypersensitivity to hormone ABA and salinity stress. Global gene expression analysis revealed AsHSP26.8a-modulated expression of heat-shock transcription factor gene, and the involvement of AsHSP26.8a in ABA-dependent and -independent as well as other stress signaling pathways. CONCLUSIONS: Our results suggest that AsHSP26.8a may negatively regulate plant response to various abiotic stresses through modulating ABA and other stress signaling pathways.

Global transcriptome analysis of alfalfa reveals six key biological processes of senescent leaves.

Leaf senescence is a complex organized developmental stage limiting the yield of crop plants, and alfalfa is an important forage crop worldwide. However, our understanding of the molecular mechanism of leaf senescence and its influence on biomass in alfalfa is still limited. In this study, RNA sequencing was utilized to identify differentially expressed genes (DEGs) in young, mature, and senescent leaves, and the functions of key genes related to leaf senescence. A total of 163,511 transcripts and 77,901 unigenes were identified from the transcriptome, and 5,133 unigenes were differentially expressed. KEGG enrichment analyses revealed that ribosome and phenylpropanoid biosynthesis pathways, and starch and sucrose metabolism pathways are involved in leaf development and senescence in alfalfa. GO enrichment analyses exhibited that six clusters of DEGs are involved in leaf morphogenesis, leaf development, leaf formation, regulation of leaf development, leaf senescence and negative regulation of the leaf senescence biological process. The WRKY and NAC families of genes mainly consist of transcription factors that are involved in the leaf senescence process. Our results offer a novel interpretation of the molecular mechanisms of leaf senescence in alfalfa.

Genome-wide analysis reveals four key transcription factors associated with cadmium stress in creeping bentgrass (Agrostis stolonifera L.).

Cadmium (Cd) toxicity seriously affects the growth and development of plants, so studies on uptake, translocation, and accumulation of Cd in plants are crucial for phytoremediation. However, the molecular mechanism of the plant response to Cd stress remains poorly understood. The main objective of this study was to reveal differentially expressed genes (DEGs) under lower (BT2_5) and higher (BT43) Cd concentration treatments in creeping bentgrass. A total of 463,184 unigenes were obtained from creeping bentgrass leaves using RNA sequencing technology. Observation of leaf tissue morphology showed that the higher Cd concentration damages leaf tissues. Four key transcription factor (TF) families, WRKY, bZIP, ERF, and MYB, are associated with Cd stress in creeping bentgrass. Our findings revealed that these four TFs play crucial roles during the creeping bentgrass response to Cd stress. This study is mainly focused on the molecular characteristics of DEGs under Cd stress using transcriptomic analysis in creeping bentgrass. These results provide novel insight into the regulatory mechanisms of respond to Cd stress and enrich information for phytoremediation.

Influence of sulfadimethoxine (SDM) and sulfamethazine (SM) on anammox bioreactors: Performance evaluation and bacterial community characterization.

The specific inhibitory effects of sulfonamides on anaerobic ammonium oxidation (anammox) process remain unknown. This study investigated the inhibitory characteristics of sulfadimethoxine (SDM) and sulfamethazine (SM) in two anammox bioreactors with NH4+-N (160 mg/L) and NO2--N (210 mg/L) in influent. Results indicate that anammox bacteria in both bioreactors adapted to low antibiotic concentrations (less than 3 mg/L). At concentrations between 5 and 7 mg/L, SDM inhibited the growth of anammox bacteria and resulted in a decrease of Candidatus Brocadia abundance from 2.57% to 0.39%. In contrast, at concentrations of 5-9 mg/L, SM inhibited the denitrification process more severely than SDM, resulting in higher accumulation of nitrite and nitrate. The purpose of this study was to elucidate the inhibitory effects of sulfonamides on the anammox process and to provide a reference for the stable operation of anammox bioreactors for the treatment of sulfonamide-containing wastewater.

Endogenous influences on anammox and sulfocompound-oxidizing autotrophic denitrification coupling system (A/SAD) and dynamic operating strategy.

The anaerobic ammonia oxidation (anammox) and sulfocompound-oxidizing autotrophic denitrification coupling system (A/SAD) was initiated in an expanded granular sludge bed (EGSB) reactor for nitrogen removal from high-strength wastewater. Owing to cooperation between anammox and partial sulfocompound-oxidation autotrophic denitrification coupling system (PSAD), the highest nitrogen removal efficiency (NRE) of 98.1% ±â€¯0.4% achieved at the optimal influent conditions of conversion efficiency of ammonium (CEA) of 55% and S2O32--S/NO3--N (S/N) of 1.4 mol mol-1. The activity of the short-cut sulfocompound-oxidizing autotrophic denitrification (SSAD) was also regulated to cope with dynamic CEA in the influent by changing the S/N, which was demonstrated to be effective in alleviating nitrite accumulation when the CEA was between 57% and 61%. Both the anammox and SAD bacteria enriched in the reactor after long-term incubation. Candidatus Brocadia and Candidatus Jettenia might be potentially contributing the most to anammox, while the Thiobacillus was the dominant taxa related to SAD.

Performance evaluation and microbial community analysis of the function and fate of ammonia in a sulfate-reducing EGSB reactor.

Ammonia is widely distributed in sulfate-reducing bioreactor dealing with sulfate wastewater, which shows potential effect on the metabolic pathway of sulfate and ammonia. This study investigates the sulfate-reducing efficiency and microbial community composition in the sulfate-reducing EGSB reactor with the increasing ammonia loading. Results indicated that, compared with low ammonia loading (166-666 mg/L), the sulfate and organic matter removal efficiencies were improved gradually with the appropriate ammonia loading (1000-2000 mg/L), which increased from 63.58 ± 3.81 to 71.08 ± 1.36% and from 66.24 ± 1.32 to 81.88 ± 1.83%, respectively. Meanwhile, with the appropriate ratio of ammonia and sulfate (1.5-3.0) and hydraulic retention time (21 h), the sulfate-reducing anaerobic ammonia oxidation (SRAO) process was occurred efficiently, inducing the accumulation of S0 (270 mg/L) and the simultaneous ammonia removal (70.83%) in EGSB reactor. Moreover, the key sulfate-reducing bacteria (SRB) (Desulfovibrio) and denitrification bacteria (Pseudomonas and Alcaligenes) were responsible for the sulfate and nitrogen removal in these phases, which accounted for 3.66-5.54 and 3.85-9.13%, respectively. However, as the ammonia loading higher than 3000 mg/L (phases 9 and 10), the sulfate-reducing efficiency was decreased to only 28.3 ± 1.26% with the ammonia removal rate of 18.4 ± 3.37% in the EGSB reactor. Meanwhile, the predominant SRB in phases 9 and 10 were Desulfomicrobium (1.22-1.99%) and Desulfocurvus (4.0-5.46%), and the denitrification bacteria accounted for only 0.88% (phase 10), indicating the low nitrogen removal rate.

Puerarin attenuates renal fibrosis by reducing oxidative stress induced-epithelial cell apoptosis via MAPK signal pathways in vivo and in vitro.

Puerarin (PR) is an isoflavonoid isolated from the root of the plant Pueraria lobata and has been widely used in traditional Chinese herbal medicine for the treatment of various diseases. Oxidative stress and epithelial cell apoptosis play important roles in the renal fibrotic process. The present study aimed to determine whether or not PR inhibits renal fibrosis by reducing oxidative stress induced-epithelial cell apoptosis. In vivo, unilateral ureteral obstruction (UUO) induced renal fibrosis, and epithelial cell apoptosis. A total of 24 mice were randomly assigned to four experimental groups: sham, UUO alone, UUO +50 mg/kg PR, and UUO +100 mg/kg PR. In vitro, 200 µM hydrogen peroxide (H2O2) induced epithelial cell apoptosis. The experiments were dived into four groups: control, H2O2 alone, H2O2+50 µM PR, and H2O2+100 µM PR. Tubular injury was measured in the renal cortex of the mice through periodic acid-Schiff (PAS) staining, and the extracellular matrix (ECM) was measured through Sirius red (SR), immunohistochemistry (IHC) staining, and Western blot. Renal epithelial cell apoptosis was measured through terminal deoxynucleotidyl transferase-mediated dUTP Nick-End labeling (TUNEL), flow cytometry (FCM), and Hoechst assays. The protein expression of NOX4, caspase3, ERK, P38, and JNK was assessed through Western blot. PAS staining showed that PR decreased renal tubular injury in UUO mice. SR and IHC staining demonstrated that PR decreased the accumulation of ECM. PR treatment significantly inhibited epithelial cell apoptosis according to the results of TUNEL, FCM, Hoechst, and Western blot. Furthermore, NOX4 increased in UUO mice and decreased with PR treatment. H2O2-derived oxidative stress activated epithelial apoptosis and mitogen-activated protein kinases (MAPK), and PR treatment significantly reversed it. These results suggest that PR treatment ameliorates renal fibrosis by inhibiting oxidative stress induced-epithelial cell apoptosis through MAPK signaling.

Slit2 ameliorates renal inflammation and fibrosis after hypoxia-and lipopolysaccharide-induced epithelial cells injury in vitro.

Hypoxic acute kidney injury (AKI) is often incompletely repaired and leads to chronic kidney disease (CKD), which is characterized by tubulointerstitial inflammation and fibrosis. The Slit2 family of secreted glycoproteins is expressed in the kidney, it has been shown to exert an anti-inflammatory activity and prevent ischemic renal injury in vivo. However, whether Slit2 reduces renal fibrosis and inflammation after hypoxic and inflammatory epithelial cells injury in vitro remains unknown. In this study, we aimed to evaluate whether Slit2 ameliorated fibrosis and inflammation in two renal epithelial cells line challenged with hypoxia and lipopolysaccharide (LPS). Renal epithelial cells were treated with hypoxia and LPS to induce cell injury. Hoechst staining and Western blot analysis was conducted to examine epithelial cells injury. Immunofluorescence staining and Western blot analysis was performed to evaluate tubulointerstitial fibrosis. Real-time polymerase chain reaction (PCR) tested the inflammatory factor interleukin (IL)-1ß and tumor necrosis factor (TNF)-α, and Western blot analysis determined the hypoxia-inducible factor (HIF)-1α, Toll-like receptor 4 (TLR4) and nuclear factor (NF)-κB. Results revealed that hypoxia induced epithelial cells apoptosis, inflammatory factor IL-1ß and TNF-α release and tubulointerstitial fibrosis. LPS could exacerbate hypoxia -induced epithelial cells apoptosis, IL-1ß and TNF-α release and fibrosis. Slit2 reduced the expression of fibronectin, the rate of epithelial cell apoptosis, and the expression of inflammatory factor. Slit2 could also inhibit the expression of TLR4 and NF-κB, but not the expression of HIF-1α. Therefore, Slit2 attenuated inflammation and fibrosis after LPS- and hypoxia-induced epithelial cells injury via the TLR4/NF-κB signaling pathway, but not depending on the HIF-1α signaling pathway.

Effect of Obesity on Outcomes of Percutaneous Nephrolithotomy in Renal Stone Management: A Systematic Review and Meta-Analysis.

BACKGROUND: Percutaneous nephrolithotomy (PCNL) has been widely used to treat renal stones. The application of PCNL in obese patients results in the emergence of a number of challenges. This study compared the effect of obesity on the outcomes of PCNL in kidney stone treatment. METHODS: Eligible studies were searched in PubMed, Web of Science, and Cochrane Library databases. Data were analyzed using RevMan statistical software, weighted mean differences, ORs, and 95% CIs were calculated. RESULTS: Seven studies involving 2,720 normal-weight, 1,686 obese, and 286 super-obese individuals were included in this meta-analysis. A pooled analysis of safety revealed that no obvious differences in terms of complication rates after treatment existed between obese and normal-weight individuals (OR 0.97, 95% CI 0.80-1.16, p = 0.73), and between super-obese and normal-weight individuals (OR 0.88, 95% CI 0.61-1.27, p = 0.49). A pooled analysis of effectiveness revealed that no obvious difference in terms of stone-free rate after treatment existed between obese and normal-weight individuals (OR 0.98, 95% CI 0.84-1.15, p = 0.79), and between super-obese and normal-weight individuals (OR 1.20, 95% CI 0.88-1.63, p = 0.25). Moreover, no obvious differences in terms of length of hospital stay after treatment existed between super-obese and normal-weight individuals (95% CI -0.15 to 0.37, p = 0.39). Additionally, no obvious differences in terms of operation time existed between obese and normal-weight individuals (95% CI -3.36 to 1.17, p = 0.34). However, the operation time was longer among super-obese individuals than among normal-weight individuals (95% CI -22.64 to -1.40, p = 0.03), and the length of hospital stay was shorter among obese patients than among normal-weight patients (95% CI 0.04-0.34, p = 0.01). No publication bias was observed in this work. CONCLUSION: The PCNL performed in normal-weight, obese, and super-obese individuals for kidney stone treatment showed similar outcomes, except that operation time was longer among super-obese individuals and the hospital stay was shorter in obese individuals than in other groups. Thus, PCNL is a safe and efficacious treatment for renal stones in patients of all sizes.

Astragaloside IV from Astragalus membranaceus ameliorates renal interstitial fibrosis by inhibiting inflammation via TLR4/NF-кB in vivo and in vitro.

Renal fibrosis is characterized by infiltration of inflammatory cells, activation and proliferation of fibroblasts, and accumulation of extracellular matrix (ECM). Astragalus membranaceus (AM) is traditional Chinese medicine and has a range of pharmacological effects. Astragaloside IV (As IV) is the main compound of AM and has anti-inflammation activities. Whether As IV ameliorates renal interstitial fibrosis by inhibiting inflammation remains unknown. Accordingly, this study investigated the ameliorating effect of As IV on renal fibrosis. Renal fibrosis was induced in vivo using the unilateral ureteral obstruction (UUO) model. UUO mice were administered intragastrically with As IV (20 and 40mg/kg/day). After a week, ECM including fibronectin and collagen I was examined by Immunohistochemistry and Western blot, inflammatory cells (CD68 and CD3) were detected by Immunohistochemistry, the release of inflammatory cytokines (tumor necrosis factor-α and interleukin-1ß) was inspected by polymerase chain reaction, and signaling pathway was determined by Western blot. In vitro, 100ng/ml lipopolysaccharide (LPS) stimulated epithelial cells to construct the inflammatory model; these cells were treated by As IV (10 and 20µM) with or without TAK-242 (1µM) for 48h. The released inflammatory cytokines were assayed by enzyme-linked immunosorbent assay, and signaling pathway was evaluated by Western blot. As IV decreased accumulation of ECM and infiltration of inflammatory cells in UUO-induced renal fibrosis. Furthermore, As IV markedly attenuated pro-inflammatory cytokines in UUO mouse and LPS-induced epithelial cells. As IV also inhibited the TLR4 and nuclear factor (NF)-кB signaling pathway in vivo and vitro. These results demonstrate that As IV protects against the progression of renal fibrosis by inhibiting inflammation via the TLR4/NF-кB signaling pathway.