Zonisamide attenuates pressure overload-induced myocardial hypertrophy in mice through proteasome inhibition.

Myocardial hypertrophy is a pathological thickening of the myocardium which ultimately results in heart failure. We previously reported that zonisamide, an antiepileptic drug, attenuated pressure overload-caused myocardial hypertrophy and diabetic cardiomyopathy in murine models. In addition, we have found that the inhibition of proteasome activates glycogen synthesis kinase 3 (GSK-3) thus alleviates myocardial hypertrophy, which is an important anti-hypertrophic strategy. In this study, we investigated whether zonisamide prevented pressure overload-caused myocardial hypertrophy through suppressing proteasome. Pressure overload-caused myocardial hypertrophy was induced in mice by trans-aortic constriction (TAC) surgery. Two days after the surgery, the mice were administered zonisamide (10, 20, 40 mg·kg-1·d-1, i.g.) for four weeks. We showed that zonisamide administration significantly mitigated impaired cardiac function. Furthermore, zonisamide administration significantly inhibited proteasome activity as well as the expression levels of proteasome subunit beta types (PSMB) of the 20 S proteasome (PSMB1, PSMB2 and PSMB5) and proteasome-regulated particles (RPT) of the 19 S proteasome (RPT1, RPT4) in heart tissues of TAC mice. In primary neonatal rat cardiomyocytes (NRCMs), zonisamide (0.3 µM) prevented myocardial hypertrophy triggered by angiotensin II (Ang II), and significantly inhibited proteasome activity, proteasome subunits and proteasome-regulated particles. In Ang II-treated NRCMs, we found that 18α-glycyrrhetinic acid (18α-GA, 2 mg/ml), a proteasome inducer, eliminated the protective effects of zonisamide against myocardial hypertrophy and proteasome. Moreover, zonisamide treatment activated GSK-3 through inhibiting the phosphorylated AKT (protein kinase B, PKB) and phosphorylated liver kinase B1/AMP-activated protein kinase (LKB1/AMPKα), the upstream of GSK-3. Zonisamide treatment also inhibited GSK-3's downstream signaling proteins, including extracellular signal-regulated kinase (ERK) and GATA binding protein 4 (GATA4), both being the hypertrophic factors. Collectively, this study highlights the potential of zonisamide as a new therapeutic agent for myocardial hypertrophy, as it shows potent anti-hypertrophic potential through the suppression of proteasome.

Design, synthesis, and hypoxia-inducible factor-1α inhibitory activity evaluation of panaxadiol derivatives containing a thiazole moiety.

The hypoxia-inducible factor-1α (HIF-1α) pathway has been implicated in tumor angiogenesis, growth, and metastasis. Therefore, the inhibition of this pathway is an important therapeutic target for cancer. Here, three series of panaxadiol derivatives containing a thiazole moiety (5a-l, 7a-i, and 9a-i) were synthesized and evaluated for HIF-1α inhibitory activity using a Hep3B cell-based luciferase reporter assay. Compounds 5d and 7b showed the strongest inhibitory activity with IC50 values of 17.37 and 6.42 µM, respectively, and did not show any significant cytotoxicity. Western blot assay results indicated that these two compounds exhibited more potent inhibition, compared with panaxadiol, of the expression of HIF-1α protein in Hep3B cells at a concentration of 50 µM. Molecular docking experiments were also performed to investigate the structure-activity relationship.

[Silk fibroin/collagen composite hydrogels with different matrix stiffness influence the growth and phenotype of human mammary epithelial cells].

Extracellular matrix (ECM) stiffness is closely related to the physiological and pathological states of breast tissue. The current study was aimed to investigate the effect of silk fibroin/collagen composite hydrogels with adjustable matrix stiffness on the growth and phenotype of normal breast epithelial cells. In this study, the enzymatic reaction of horseradish peroxidase (HRP) with hydrogen peroxide (H2O2) was used to change the degree of cross-linking of the silk fibroin solution. The rotational rheometer was used to characterize the composite hydrogel's biomechanical properties. Human normal mammary epithelial cell line MCF-10A were inoculated into composite hydrogels with various stiffness (19.10-4 932.36 Pa) to construct a three dimensional (3D) culture system of mammary epithelial cells. The CCK-8 assay was applied to detect the cell proliferation rate and active states in each group. Hematoxylin-Eosin (HE) staining and whole-mount magenta staining were used for histological evaluation of cell morphology and distribution. The results showed that with the increase of matrix stiffness, MCF-10A cells exhibited inhibited proliferation rate, decreased formation of acinus structures and increased branching structures. Meanwhile, with the increase of matrix stiffness, the polarity of MCF-10A cells was impeded. And the increase of matrix stiffness up-regulated the expression levels of mmp-2, mmp-3, and mmp-9 in MCF-10A cells. Among the genes related to epithelial-mesenchymal transition (EMT), the expression level of the epithelial marker gene E-cadherin was significantly down-regulated, while the interstitial cell marker gene Vimentin was up-regulated, and the expression levels of Snail, Wnt5b and Integrin ß1 in the Wnt pathway were up-regulated. These results suggest that the silk fibroin/collagen composite hydrogels with adjustable matrix stiffness regulates the proliferation and the phenotype of MCF-10A cells. The effects of increased matrix stiffness may be closely related to the changes of the polar structures and function of MCF-10A cells, as well as the occurrence of ECM-remodeling and EMT.

[Identification of Hub Genes for Ovarian Cancer Stem Cell Properties with Weighted Gene Co-expression Network Analysis].

OBJECTIVE: To investigate the significance of stemness-related genes in the diagnosis and treatment of ovarian cancer. METHODS: Key modules and genes were identified with weighted gene co-expression network analysis (WGCNA). The signal pathways of high expression of key genes were analyzed by gene set enrichment analysis (GSEA) and single cell sequencing data. The chemosensitivity of ovarian cancer to chemotherapy drugs was estimated with pRRophetic. Flow cytometry was used to examine the expression of CD44 +CD117 +in SKOV3 cells and cancer stem cells. The expression of key genes in ovarian cancer stem cells was confirmed by qRT-PCR. The core genes were identified by GeneMANIA analysis. RESULTS: According to the WGCNA results, 15 key genes were identified at the transcription level, all being highly expressed in many kinds of tumors. They were involved in the cell cycle, DNA repair, E2 target and G2M checkpoint pathway, and had significant correlation with chemosensitivity. The proportion of CD44 + CD117 + cells in SKOV3 cells and ovarian cancer stem cells were (1.20±0.34)% and (37.17±1.80)% respectively, with statistically significant difference ( P<0.05). qRT-PCR confirmed that seven key genes ( BUB1, CDC20, CCNB2, DLGAP5, KIF4 A, NEK2, NUSAP1) in the WGCNA results were highly expressed in ovarian cancer stem cells, and BUB1 might have played a core role. CONCLUSION: Seven hub genes, especially BUB1, were identified by constructing gene co-expression network, which may become potential biomarkers of ovarian cancer gene.

LncRNA-ANRIL inhibits cell senescence of vascular smooth muscle cells by regulating miR-181a/Sirt1.

BACKGROUND: Cardiovascular disease is one of the major threats to human life and health, and vascular aging is an important cause of its occurrence. Antisense non-coding RNA in the INK4 locus (ANRIL) is a kind of long non-coding RNA (lncRNA) that plays important roles in cell senescence. However, the role and mechanism of ANRIL in senescence of vascular smooth muscle cells (VSMCs) are unclear. METHODS: Cell viability and cell cycle were evaluated using an MTT assay and flow cytometry analysis, respectively. Senescence-associated (SA)-ß-galactosidase (gal) staining was used to determine cell senescence. Dual luciferase reporter assays were conducted to confirm the binding of ANRIL and miR-181a, as well as miR-181a and Sirt1. The expression of ANRIL, miR-181a, and Sirt1 was determined using qRT-PCR and protein levels of SA-ß-gal and p53-p21 pathway-related proteins were evaluated by Western blotting. RESULTS: ANRIL and Sirt1 were down-regulated, whereas miR-181a was up-regulated in aging VSMCs. In young and aging VSMCs, over-expression of ANRIL could down-regulate miR-181a and up-regulate Sirt1. MTT and SA-ß-gal staining assays showed that over-expression of ANRIL and inhibition of miR-181a promoted cell viability and inhibited VSMC senescence. The dual-luciferase reporter assay determined that miR-181a directly targets ANRIL and the 3'-UTR of Sirt1. Furthermore, over-expression of ANRIL inhibited cell cycle arrest and the p53-p21 pathway. CONCLUSION: ANRIL promotes cell viability and inhibits senescence in VSMCs, possibly by regulating miR-181a/Sirt1, and alleviating cell cycle arrest by inhibiting the p53-p21 pathway. This study provides novel insights for the role of ANRIL in the development of cell senescence.

Dietary Aloe vera supplementation on growth performance, some haemato-biochemical parameters and disease resistance against Streptococcus iniae in tilapia (GIFT).

This study investigated effects of dietary Aloe vera on growth performance, some haemato-biochemical parameters and disease resistance against Streptococcus iniae in tilapia (GIFT). Five groups were designed including a basal diet (control) and 100% A. vera powder incorporated in fish feed at 0.5% 1%, 2%, and 4%/kg feed, which were administered for 8 weeks. Fish fed 0.5%, 1%, and 2% A. vera supplemented diet significantly improved (p < 0.05) weight gain, absolute growth rate and specific growth rate. Feed intake significantly increased in fish fed with A. vera diet at 1% and 2%/kg feed. Feed efficiency ratio, feed conversion ratio, and hepatosomatic index were significantly enhanced in 4% A. vera supplemented fish over unsupplemented ones (p < 0.05). Several haemato-biochemical indices were examined before and after fish were challenged with S. iniae pathogen containing 7.7 × 10(6) CFU cells mL(-1). A. vera supplemented fish showed a significant increase (p < 0.05) in red blood cells, hematocrits (Hb), hemoglobin (Hb), white blood cells (WBC), neutrophils, monocytes, eosinophils, serum total protein, glucose and cortisol after challenge when compared to unsupplemented ones. Meanwhile, 4% A. vera supplemented fish showed a decrease (p < 0.05) in RBC, Hb, Ht, WBC, and mean corpuscular hemoglobin (MCH) after challenge compared to unsupplemented ones and other supplemented ones. In addition, lower mean corpuscular volume values (MCV) (p < 0.05) were observed in fish fed with A. vera diet at 2% and 4% A. vera/kg feed than those fed unsupplemented diet. Unchallenged fish fed 0.5%, 1%, and 2% A. vera showed significantly higher values (p < 0.05) of mean corpuscular hemoglobin concentration (MCHC) than those fed unsupplemented diet and 4% A. vera supplemented diet. There was a significant increase (p < 0.05) in the neutrophil to lymphocyte ratio (N/L) within experimental groups after challenge; N/L ratio in A. vera unsupplemented fish and those supplemented with A. vera diet at 1%/kg feed increased significantly (p < 0.05) throughout challenge period; while those fed 4% A. vera supplemented diet maintained higher values at all experimental stages among groups. There was a significant correlation (p < 0.05, r = 0.53) between N/L ratio and glucose concentration, 96 h after challenge. Aloe had no significant effect (p > 0.05) on the survival of the fish when compared to the control; no mortality was recorded in challenge trial. Overall, our results indicated that dietary aloe supplementation could improve growth, feed utilization, and haemato-biochemical parameters of cultured tilapia.

Dietary Aloe vera improves plasma lipid profile, antioxidant, and hepatoprotective enzyme activities in GIFT-tilapia (Oreochromis niloticus) after Streptococcus iniae challenge.

The current study investigated the effects of dietary Aloe vera on plasma lipid profile status, antioxidant, and hepatoprotective enzyme activities of GIFT-tilapia juveniles under Streptococcus iniae challenge. Five dietary groups were designed including a control and 100 % Aloe powder incorporated into a tilapia feed at 0.5, 1, 2, and 4 %/kg feed, which were administered for 8 weeks. Fish fed dietary Aloe at 4 %/kg feed significantly reduced in total cholesterol, while triacylglycerol reduced (P < 0.05) in those fed 0.5, 2, and 4 % Aloe/kg feed compared to unsupplemented ones. High-density lipoprotein was significantly elevated in fish fed 0.5 and 1 % Aloe/kg feed compared to unsupplemented ones, and no significant changes (P > 0.05) were noted in low-density lipoprotein among test groups. Furthermore, high activities of superoxide dismutase, catalase, and glutathione peroxide in liver tissues were observed in Aloe-supplemented fish compared to unsupplemented ones, before and after S. iniae challenge (7.7 × 10(6) CFU cells/mL). Variations were also noted in malondialdehyde activity throughout the trial, but no significant difference (P > 0.05) was observed between groups. Meanwhile, Aloe-supplemented fish reduced serum aspartate and alanine aminotransferase (AST and ALT) activities before and after challenge. Based on the second-order polynomial regression analysis, dietary Aloe inclusion levels less than or equal to 1.88, 1.86, and 2.79 %/kg feed were determined to be suitable in improving plasma lipid profile status, antioxidant, and hepatoprotective enzyme activities in GIFT-tilapia in this study, respectively. Thus, A. vera extracts may be recommended as a tilapia feed supplement to enhance fish antioxidant and hepatoprotective capacities, especially during disease outbreaks.

Ependyma in Neurodegenerative Diseases, Radiation-Induced Brain Injury and as a Therapeutic Target for Neurotrophic Factors.

The neuron loss caused by the progressive damage to the nervous system is proposed to be the main pathogenesis of neurodegenerative diseases. Ependyma is a layer of ciliated ependymal cells that participates in the formation of the brain-cerebrospinal fluid barrier (BCB). It functions to promotes the circulation of cerebrospinal fluid (CSF) and the material exchange between CSF and brain interstitial fluid. Radiation-induced brain injury (RIBI) shows obvious impairments of the blood-brain barrier (BBB). In the neuroinflammatory processes after acute brain injury, a large amount of complement proteins and infiltrated immune cells are circulated in the CSF to resist brain damage and promote substance exchange through the BCB. However, as the protective barrier lining the brain ventricles, the ependyma is extremely vulnerable to cytotoxic and cytolytic immune responses. When the ependyma is damaged, the integrity of BCB is destroyed, and the CSF flow and material exchange is affected, leading to brain microenvironment imbalance, which plays a vital role in the pathogenesis of neurodegenerative diseases. Epidermal growth factor (EGF) and other neurotrophic factors promote the differentiation and maturation of ependymal cells to maintain the integrity of the ependyma and the activity of ependymal cilia, and may have therapeutic potential in restoring the homeostasis of the brain microenvironment after RIBI or during the pathogenesis of neurodegenerative diseases.

[Effects of Different Carbon Inputs on Soil Stoichiometry in Tianshan Mountains].

Soil C, N, and P elements are important components of the forest ecosystem. Studying the influence of exogenous carbon input change on the stoichiometry of the forest soil can reveal the element recycling process and the balanced feedback mechanism of the forest ecosystem. In this study, using the research object of a spruce forest in Tianshan Mountain, the short-term effect of exogenous carbon input on soil C, N, and P in the soil was analyzed through Detritus Input and Removal Treatment (DIRT), and then the interrelationship between soil stoichiometry and other soil physicochemical factors under different treatments was discussed. The results showed that:① the soil C, N, and P contents in most soil layers were the highest double litter (DL) treatment, soil ω(C) by soil depth from shallow to deep was 168.92, 119.88, 103.33, and 64.23 g·kg-1; soil ω(N) was 10.60, 9.32, 8.78, and 8.07 g·kg-1; soil ω(P) was 0.50, 0.45, 0.37, and 0.36 g·kg-1; in the no input (NI) treatment, soil ω(C) by soil depth from shallow to deep was 104.56, 89.24, 48.08, and 43.96 g·kg-1; soil ω(N) was 6.83, 2.60, 2.63, and 2.22 g·kg-1; soil ω(P) was 0.40, 0.34, 0.32, and 0.22 g·kg-1; and a decreased trend was shown with the deepening of the soil layer. Except in the NI treatment, C:N was 0-10 cm and significantly higher than that in other soils (P<0.05), NL soil C:P at 30-50 cm was significantly higher than that in other soils, and NI soil N:P was 0-10 cm and significantly higher than that in other soils (P<0.05). ② Microbial carbon, nitrogen, and phosphorus were significantly higher from 0-10 cm than that in other soil layers (P<0.05). ③ Redundancy analysis results showed that soluble organic carbon and microbial nitrogen at different carbon input levels were important factors affecting the stoichiometric characteristics of soil C, N, and P.

A bacteriocyte symbiont determines whitefly sex ratio by regulating mitochondrial function.

Nutritional symbionts influence host reproduction, but the underlying molecular mechanisms are largely unclear. We previously found that the bacteriocyte symbiont Hamiltonella impacts the sex ratio of the whitefly Bemisia tabaci. Hamiltonella synthesizes folate by cooperation with the whitefly. Folate deficiency by Hamiltonella elimination or whitefly gene silencing distorted whitefly sex ratio, and folate supplementation restored the sex ratio. Hamiltonella deficiency or gene silencing altered histone H3 lysine 9 trimethylation (H3K9me3) level, which was restored by folate supplementation. Genome-wide chromatin immunoprecipitation-seq analysis of H3K9me3 indicated mitochondrial dysfunction in symbiont-deficient whiteflies. Hamiltonella deficiency compromised mitochondrial quality of whitefly ovaries. Repressing ovary mitochondrial function led to distorted whitefly sex ratio. These findings indicate that the symbiont-derived folate regulates host histone methylation modifications, which thereby impacts ovary mitochondrial function, and finally determines host sex ratio. Our study suggests that a nutritional symbiont can regulate animal reproduction in a way that differs from reproductive manipulators.

[Sediment Pollution Characteristics and Dredging in the Nanfei River Estuary, Chaohu Lake].

To clarify the endogenous pollution characteristics of sediments in the Nanfei River estuary, Chaohu Lake, and provide a theoretical basis for the dredging works, the organic index, pollution index, and potential ecological risk were determined. The results show that the average total nitrogen (TN), total phosphorus (TP), and organic matter (OM) content of the sediment was 1461 mg·kg-1, 438 mg·kg-1, and 1.77%, respectively, showing enrichment in the surface layer (0-10 cm). The nutrient pollution status and TP pollution index show that the organic pollution level was moderate and the TP pollution was severe. Furthermore, the pollution risk gradually reduced with sediment depth, representing a low risk at depths below 30 cm. Static release results showed that the average release fluxes of NH4+-N and TP in the sediment were 8.04 mg·(m2·d)-1 and 0.19 mg·(m2·d)-1, respectively, and showed highest release potentials consistent with areas of sediment nutrient pollution. Except for Cr and Ni, the concentrations of six heavy metals were higher than the soil background values for Anhui Province, and the concentrations of Hg and Cd far exceeded the standards. According to the assessment of potential ecological risk from heavy metals, the 0-20 cm sediments present a high level of risk and sediments below 30 cm have a low level of risk. Heavy metal leaching toxicity indicated that the risk of heavy metal release after dredging is low and non-hazardous. These results were used to determine the key dredging area (3.93 km2) and depth (30 cm) for the Nanfei River estuary, providing an important basis for future dredging activities.

Vaccarin enhances intestinal barrier function in type 2 diabetic mice.

AIMS: Hyperglycemia and insulin resistance drive intestinal barrier dysfunction in type 2 diabetes (T2DM). Vaccarin, the main active component in the semen of traditional Chinese medicine Vaccaria has a definite effect on T2DM mice. The purpose of this study was to investigate whether vaccarin can enhance the intestinal barrier function in T2DM. MAIN METHODS: The T2DM mice model was established by streptozocin and high-fat diet. Vaccarin at a dose of 1 mg/kg/day was administered. We evaluated the effects of vaccarin on gut microbiota and intestinal barrier function by 16S rRNA sequencing, Western blot, quantitative fluorescent PCR (qPCR), and morphological observation. Moreover, we constructed a single layer of the human intestinal epithelium model to determine the effect of vaccarin in vitro. RESULTS: The experimental results showed that vaccarin alleviated inflammatory mediators in serum and intestinal tissue of mice (P < 0.05), which may depend on the improvement of tight junctions and gut microbiota (P < 0.05). Activation of extracellular regulated protein kinases (Erk1/2) stimulated myosin light chain kinase (MLCK). By inhibiting ERK expression (P < 0.05), vaccarin had similar effects to ERK inhibitors. In addition, the regulation of tight junction barriers also involved the abovementioned pathways in vivo. CONCLUSION: Vaccarin could protect the intestinal barrier by inhibiting the ERK/MLCK signaling pathway and modulate the composition of the microbiota. These results suggested that vaccarin may be an effective candidate for improving intestinal barrier changes in T2DM.

Survey of antibiotic use among hospitalised children in a hospital in Northeast China over a 4-year period.

PURPOSE: We investigated the usage of antibiotic utilisation rate (AUR) among Chinese hospitalised children over a 4-year period to determine clinical characteristics. DESIGN AND METHODS: AUR, antibiotic type and antibiotic use in combination were analysed among hospitalised children of the affiliated hospital of Beihua University in Northeast China from January 2015 to December 2018. The linear prediction was used to investigate the trends of antibiotics use in combination and AUR, and autoregressive integrated moving average model was used to predict AUR in 2019. RESULTS: A total of 2,981 inpatients were admitted to the hospital during the study period. The AUR from 2015 to 2018 was 91.51%, 92.67%, 91.30%, and 93.00%, respectively. AUR was associated with season (p < .01), the peak period was found to be from November to January in 2019 (R2 = 0.802). The etiological delivery rate had increased (p < .01). Pneumonia was the main disease in children for which they received antibiotics. The combination of multidrug (≥3 agents used) had increased (p < .01) over the study period. PRACTICE IMPLICATIONS: AUR was stable among hospitalised children in a hospital. Cephalosporin and macrolide antibiotics were the main antibiotic types, and the combination of multidrug had increased, more attention should be paid to the use of antibiotics in hospitalised children.

Construction of a novel vector expressing Survivin-shRNA and fusion suicide gene yCDglyTK and its application in inhibiting proliferation and migration of colon cancer cells.

Despite progress achieved in cancer chemotherapy in recent decades, adverse effects remain a limiting factor for a number of patients with colorectal cancer, suggesting the requirement for novel therapeutic strategies. Gene therapy appears to be a promising strategy for treating cancer. The present study aimed to investigate the anti-tumor effect of a combined gene therapy, using Survivin downregulation by RNAi and a fusion suicide gene yCDglyTK therapy system. A triple-gene vector expressing Survivin-targeted small hairpin RNA (Survivin-shRNA) and fusion suicide gene yCDglyTK was constructed, and administered to HCT116 cells. Survivin expression decreased significantly and yCDglyTK fusion gene expression was confirmed by both reverse transcription-quantitative polymerase chain reaction and western blot analysis. Introduction of Survivin-shRNA into yCDglyTK/prodrug system eradicated colon cancer cells and induced apoptosis more effectively. Furthermore, this therapeutic system is able to inhibit the migration of HCT116 cells. These results indicate that the recombinant plasmid may serve as a novel gene therapy approach to treat colorectal carcinoma.