Association of phthalate exposure with type 2 diabetes and the mediating effect of oxidative stress: A case-control and computational toxicology study.

Phthalic acid esters (PAEs) are widely used as plasticizers and have been suggested to engender adverse effects on glucose metabolism. However, epidemiological data regarding the PAE mixture on type 2 diabetes (T2DM), as well as the mediating role of oxidative stress are scarce. This case-control study enrolled 206 T2DM cases and 206 matched controls in Guangdong Province, southern China. The concentrations of eleven phthalate metabolites (mPAEs) and the oxidative stress biomarker 8-hydroxy-2'-deoxyguanosine (8-OHdG) in urine were determined. Additionally, biomarkers of T2DM in paired serum were measured to assess glycemic status and levels of insulin resistance. Significantly positive associations were observed for mono-(2-ethylhexyl) phthalate (MEHP) and Mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP) with T2DM (P < 0.001). Restricted cubic spline modeling revealed a non-linear dose-response relationship between MEHHP and T2DM (Pnon-linear = 0.001). The Bayesian kernel machine regression and quantile g-computation analyses demonstrated a significant positive joint effect of PAE exposure on T2DM risk, with MEHHP being the most significant contributor. The mediation analysis revealed marginal evidence that oxidative stress mediated the association between the mPAEs mixture and T2DM, while 8-OHdG respectively mediated 26.88 % and 12.24 % of MEHP and MEHHP on T2DM risk individually (Pmediation < 0.05). Di(2-ethylhexyl) phthalate (DEHP, the parent compound for MEHP and MEHHP) was used to further examine the potential molecular mechanisms by in silico analysis. Oxidative stress may be crucial in the link between DEHP and T2DM, particularly in the reactive oxygen species metabolic process and glucose import/metabolism. Molecular simulation docking experiments further demonstrated the core role of Peroxisome Proliferator Activated Receptor alpha (PPARα) among the DEHP-induced T2DM. These findings suggest that PAE exposure can alter oxidative stress via PPARα, thereby increasing T2DM risk.

TiO2-PAA-SiO2 pearl chain blend modified polyvinylidene fluoride ultrafiltration membrane with excellent oil-water separation, anti fouling performance and durability.

In this work, a new type of composite nanoparticles, 'pearl chain', was developed by linking titanium dioxide and silicon dioxide by polyacrylic acid polymer chains, and the prepared TiO2-PAA-SiO2 composite nanoparticles were analysed by SEM, Fourier transform infrared spectroscopy, x-ray photoelectron spectroscopy and thermogravimetric analysis, zeta potential, x-ray diffraction, etc. The success of this work was verified by the successful linking of TiO2-PAA-SiO2 composite nanoparticles.TiO2-PAA-SiO2 composite nanoparticles were analysed to verify the successful attachment of pearl chains. The obtained TiO2-PAA-SiO2 were subsequently blended in different ratios to prepare polyvinylidene fluoride (PVDF) ultrafiltration membranes. The membrane performance was tested by porosity and water contact angle measurements, scanning electron microscopy, as well as experiments using bovine serum proteins and MTBE interception. The results showed that when a certain amount of TiO2-PAA-SiO2 was added, the surface wettability, porosity and permeability of the prepared modified composite membranes were significantly improved, and the BSA adsorption rate was increased from 71.59% to 80.86%, and the retention rate of MTBE was increased by 77%, in addition to showing a better anti-pollution effect (FRR: 91.07%). It was finally concluded that the prepared membranes embedded with 1.0 wt.% TiO2-PAA-SiO2 nanofillers showed good overall filtration performance, better contamination resistance and remarkable durability. The present work successfully demonstrated the feasibility of using polyacrylic acid chemical chains to connect nanoparticles with different functions to prevent particle loss and substantially enhance membrane performance, which is valuable for bridging connection of composite nanoparticles and exploring the development of high-performance ultrafiltration membranes.

Study on the Thermophysical Properties of 80% 10B Enrichment of B4C.

In this paper, a specific type of Boron Carbide (B4C) with a high enrichment of 80 ± 0.3 at% 10B was prepared as an absorbing material for control rods in nuclear reactors. The enrichment of 10B was achieved using a chemical exchange method, followed by obtaining boron carbide powder through a carbothermal reduction method. Finally, B4C with a high enrichment of 68.3~74.2% theoretical density was obtained using a hot-pressed sintering process. This study focused on investigating the basic out-of-pile thermophysical properties of the high enrichment B4C compared to natural B4C reference pellets under non-irradiated conditions. These properties included the thermal expansion coefficient, thermal conductivity, emissivity, elastic limit, elastic modulus, and Poisson's ratio. The research results indicate that the enriched B4C pellet exhibits good thermal stability and meets the technical requirements for mechanical capability. It was observed that porosity plays a significant role in determining the out-of-pile mechanical capability of B4C, with higher porosity samples having a lower thermal conductivity, elastic-plastic limit, and elastic modulus. In short, all the technical indexes studied meet the requirements of nuclear-grade Boron Carbide pellets for Pressurized Water Reactors.

Cervical lymph node metastasis of bladder cancer: a case report and review of literature.

OBJECTIVE: To report an extremely rare case of bladder cancer patient with cervical lymph nodes, abdominal lymph nodes, and bone metastases at the same time. METHODS AND RESULTS: The case was investigated by follow-up and immunohistochemistry was used in the pathological part. RESULT: The patient was diagnosed with bladder cancer (high-grade urothelial metastatic epithelial cell carcinoma) by pathology and immunohistochemistry after transurethral resection of bladder tumor (TURBT) and metastatic bladder cancer by pathology and immunohistochemistry after cervical lymph node aspiration due to neck lymph node enlargement 1 year later, and a CT of the chest and abdomen suggested that the patient also had abdominal lymph node and bone metastases.At the 2.5-year regular chemotherapy follow-up, the patient showed that the abdominal lymph node metastasis disappeared, the cervical lymph node fusion shrank, and the bone metastasis still existed. CONCLUSION: 1. Regular postoperative review is particularly important; 2.For patients with UCB who undergo TURBT, a effective regular perfusion program should be performed throughout the postoperative period; 3. For patients with postoperative metastatic symptoms of UCB, Complex treatment has a positive effect on patient prognosis; 4.The presence of enlarged head and neck lymph nodes in patients with bladder cancer should also be considered as metastatic of UCB.

Association of urinary iodine concentration with prediabetes/diabetes in adults: Analysis of the NHANES 2005-2016.

BACKGROUND & AIMS: Iodine is required for synthesizing thyroid hormones and regulating human metabolism. Iodine deficiency can lead to thyroid function abnormalities, which are closely associated with disturbances in glucose-insulin homeostasis. Research on the relationship between iodine and diabetes/prediabetes in adults was sparse and inconsistent. We assessed trends in urinary iodine concentration (UIC) and diabetes/prediabetes prevalence and focused on the association between iodine and diabetes/prediabetes among U.S. adults. METHODS: We analyzed the National Health and Nutrition Examination Survey (NHANES) data from the 2005-2016 cycles. Linear regression was employed to evaluate UIC and prediabetes/diabetes prevalence trends over time. Both multiple logistic regression and restricted cubic splines (RCS) were performed to evaluate the association of UIC with diabetes/prediabetes. RESULTS: A distinctly declining trend in median UIC and a significant increase in diabetes prevalence in U.S. adults from 2005 to 2016 were observed. The fourth quartile of UIC was associated with a 30% lower risk for prediabetes, compared with the first quartile (OR=0.70, 95% CI: 0.56-0.86, Ptrend=0.001). However, UIC was not significantly associated with the prevalence of diabetes. The RCS model suggested a significant nonlinear relationship between UIC and the risk of diabetes (P for nonlinearity =0.0147). Stratification analysis showed that the negative associations of UIC with the risk of prediabetes were more pronounced in participants who were men, aged 46-65, overweight, light alcohol drinkers, and nonactive smokers. CONCLUSIONS: Overall, the adults' median UIC in the U.S. population was a declining trend. However, diabetes prevalence increased significantly from 2005 to 2016. Higher UIC was associated with a lower risk of prediabetes.

Separation of oil vapour by polyether block amide composite membrane modified with porous materials.

The ability of membranes to separate oil vapour is affected by their permeance and selectivity. This study modifies polyether block amide (PEBA) composite membranes with a microporous zeolite, Silicalite-1, or a mesoporous zeolite, MCM-41. The results show that when PEBA composite membranes are modified with these zeolites, the selective layer of the composite membrane is coated more thinly, resulting in a higher flux of organic gas. Silicalite-1 increases the hydrophobicity of the membrane, which facilitates the adsorption of organic vapour on the membrane surface, thus improving the membrane selectivity. In the separation of oil vapour, both modified membranes can effectively increase the gas permeabilities and selectivities. The main mechanism governing gas transport in the MCM-41-modified membrane is Knudsen diffusion, so the selectivity for small molecules is improved more significantly. By contrast, the dissolution-diffusion mechanism is dominant in the Silicalite-1-modified membranes, which considerably increases the selectivity for large molecules.

Axial Crystal Growth Evolution and Crystallization Characteristics of Bi-Continuous Polyamide 66 Membranes Prepared via the Cold Non-Solvent-Induced Phase Separation Technique.

Polyamide 66 microporous membranes were prepared by cold non-solvent-induced phase separation using polyamide 66-formic acid-propylene carbonate as a ternary membrane-forming system. The formed membranes exhibited a special bicontinuous structure consisting of interglued spherical crystals or interlocked bundles of microcrystalline aggregates. The variation of the microporous structure under the influence of preparation conditions, solvent, aging time, and polymer concentration affects the comprehensive performance of the membranes. For example, the cold-induced operation and the use of different membrane-forming solvents contributed to the crystallization of polyamide 66, resulting in an increased contact angle of polyamide 66 membranes, obtaining a high resistance to contamination of up to 73.5%. Moreover, the formed membranes still have high mechanical strength.

Graphene Oxide Modified Polyamide 66 Ultrafiltration Membranes with Enhanced Anti-Fouling Performance.

Improving the contamination resistance of membranes is one of the most effective ways to address the short service life of membranes. While preparing the membrane system structure, doping nanoparticles into the polymer matrix is beneficial to the preparation of high-performance membranes. To develop a new structure for membrane contamination protection, in this study, a novel asymmetric polyamide 66 composite ultrafiltration (UF) membrane was fabricated by incorporating different masses (ranging from zero to 0.5 wt.%) of graphene oxide (GO) into the polyamide 66 microporous substrate, using formic acid and propylene carbonate as solvents. The effects of GO doping on the morphology, microporous structure and surface of ultrafiltration membranes were investigated by atomic force microscopy (AFM), scanning electron microscopy (SEM), integrated thermal analysis (DSC) and contact angle (CA). In addition, pure water flux, bovine serum albumin (BSA) rejection and contamination resistance were measured to evaluate the filtration performance of different membranes. The overall performance of all the modified membranes was improved compared to pure membranes. The results of contact angle and permeation experiments showed that the addition of GO improved the hydrophilicity of the membrane, but reduced the permeability of the membrane. The minimum flux was only 3.5 L/m2·h, but the rejection rate was 92.5%. Most noteworthy was the fact that GO further enhanced the anti-pollution performance of the membranes and achieved a remarkable performance of 91.32% when the GO content was 0.5 wt.%, which was 1.36 times higher than that of the pure membrane. Therefore, optimal performance was achieved. Furthermore, the UF membrane made of composite substrate offers a promising solution for the development of long-life ultrafiltration membranes with better stability, high-cost efficiency and adequate chemical durability.

Transcriptomic Variations and Network Hubs Controlling Seed Size and Weight During Maize Seed Development.

To elucidate the mechanisms underlying seed development in maize, comprehensive RNA-seq analyses were conducted on Zhengdan1002 (ZD1002), Zhengdan958 (ZD958), and their parental lines during seven seed developmental stages. We found that gene expression levels were largely nonadditive in hybrids and that cis-only or trans × cis pattern played a large role in hybrid gene regulation during seed developmental stage. Weighted gene co-expression network (WGCNA) analysis showed that 36 modules were highly correlated (r = -0.90-0.92, p < 0.05) with kernel weight, length, and width during seed development. Forty-five transcription factors and 38 ribosomal protein genes were identified as major hub genes determining seed size/weight. We also described a network hub, Auxin Response Factor 12 of maize (ZmARF12), a member of a family of transcription factor that mediate gene expression in response to auxin, potentially links auxin signal pathways, cell division, and the size of the seeds. The ZmARF12 mutant exhibited larger seed size and higher grain weight. ZmARF12 transcription was negatively associated with cell division during seed development, which was confirmed by evaluating the yield of protoplasts that isolated from the kernels of the mutant and other inbred lines. Transient knock-down of ZmARF12 in maize plants facilitated cell expansion and division, whereas transient silencing of its potential interactor ZmIAA8 impaired cell division. ZmIAA8 expression was repressed in the ZmARF12 over-expressed protoplasts. The mutant phenotype and the genetics studies presented here illustrated evidence that ZmARF12 is a cell division repressor, and potentially determines the final seed size.

A brief review and case report of urothelial carcinoma and metachronous leiomyosarcoma of the bladder at the same anatomic region.

One patient with bladder leiomyosarcoma and urothelial carcinoma is very rare. Only 10 cases have been reported in the literature. A 70-year-old patient was admitted due to bladder tumor. Two TURBTs were performed confirming the patient was free of tumor, and pathology reported low-grade urothelial carcinoma. Three years later, a tumor was also found on the right anterolateral wall of urinary bladder and was diagnosed as leiomyosarcoma by pathological examination. Radical cystectomy was performed. With 45 months follow-up, the patient has no recurrence. Two malignancies in the same anatomic region at different time has never been reported to date.

The association of aldehydes exposure with diabetes mellitus in US population: NHANES 2013-2014.

BACKGROUND: The association of mixed aldehydes exposure with diabetes remains unclear. OBJECTIVE: We aimed to explore associations between serum aldehydes concentration and diabetes. METHODS: We analyzed associations between aldehydes and diabetes using data from 1795 participants in the National Health and Nutrition Examination Survey (NHANES) from 2013 to 2014 by multiple logistic regression models. Bayesian kernel machine regression (BKMR) was used to evaluate the combined association of serum aldehydes on prediabetes and diabetes. RESULTS: Isopentanaldehyde increased the risk of diabetes 2.09 fold (95%CI:1.05-4.16) in the highest tertile, compared to the lowest-tertile concentration after adjusting for covariates, with a p-value for trend (P-t) equal to 0.041, in females. The adjusted OR of prediabetes with a 95% CI for the highest tertile was 0.52(0.28, 0.97) for benzaldehyde in females (P-t = 0.034). We also found associations in the male group between butyraldehyde and diabetes for the second (OR:2.80, 95%CI:1.35-5.79) and third (OR:2.59, 95%CI:1.30-5.17) tertile levels (P-t = 0.010). The risk of diabetes increased 2.55 fold (95%CI: 1.26-5.16, P-t = 0.008), in subjects in the highest tertile of hexanaldehyde concentration. Other aldehydes did not show a statistically significant association with diabetes or prediabetes. The BKMR model showed a positive association of mixed aldehydes with diabetes in males, and butyraldehyde showed a significant positive trend with the highest posterior inclusion probability (PIP = 0.85). Mixed aldehydes increased female's risk from prediabetes to diabetes in which isopentanaldehyde had the highest posterior inclusion probability (PIP = 0.67). CONCLUSIONS: The mixed aldehydes might increase the risk of suffering from diabetes in males and accelerate the progression of diabetes in females, in which butyraldehyde and isopentanaldehyde play the most important roles.

Lycium barbarum polysaccharides attenuate cardiovascular oxidative stress injury by enhancing the Keap1/Nrf2 signaling pathway in exhaustive exercise rats.

Moderate exercise is beneficial to physical and mental health. When the amount of exercise and exercise intensity exceeds a certain limit and reaches the state of exhaustion, oxidative stress levels in the body increase, which can lead to oxidative stress­associated damage. Lycium barbarum polysaccharide (LBP) is one of the primary active ingredients extracted from wolfberry. Following exhausting exercise in rats, LBP supplements decrease damage to the myocardium and blood vessels, indicating that LBP exerts a protective effect on the cardiovascular system. The Kelch­like ECH­associated protein 1 (Keap1)/NF­E2­related factor 2 (Nrf2) anti­oxidative stress signaling pathway improves total oxidizing ability; anti­apoptosis and other aspects serve a vital role. In the present study, LBP intervention was performed in vivo and in vitro to observe its effect on the Keap1/Nrf2 pathway and oxidative stress­associated indicators in order to clarify its protective mechanism. For the in vivo experiments, 60 male Sprague­Dawley rats were randomly divided into normal control and aerobic, exhaustive and exhaustive exercise + LBP (200 mg/kg/day) groups. For the in vitro experiments, a rat thoracic aortic endothelial cell (RTAEC) oxidative stress model was established using angiotensin II (AngII) and divided into blank control, LBP (3,200 µg/ml), AngII (1x10­4 mol/l) and AngII + LBP groups. For in vitro experiments, small interfering (si)RNA (50 nmol) was used to transfect RTAEC and induce gene silencing of Nrf2. ELISA, hematoxylin and eosin staining, TUNEL, immunofluorescence, western blotting, immunohistochemistry and reverse transcription­quantitative PCR were used to evaluate and verify the effect of LBP on oxidative stress indicators and the expression of Keap1/Nrf2 antioxidative stress signaling pathway. The in vivo experiments showed that LBP decreased the expression of serum malondialdehyde (MDA) and AngII, as well as apoptosis of blood vessels and cardiomyocytes and expression of TNF­α in rats following exhaustive exercise. Meanwhile, LBP enhanced expression of the Keap1/Nrf2 signaling pathway and downstream associated protein glutamyl­cysteine synthetase catalytic subunit (GCLC), quinone oxidoreductase 1 (NQO1) and glutamate­cysteine ligase modified subunit (GCLM) in the thoracic aorta and myocardium of rats following exhaustive exercise. In RTAEC in vitro, LBP decreased the expression of MDA and TNF­α in the supernatant, promoted the nuclear translocation of Nrf2 and increased expression levels of GCLC, NQO1 and GCLM. Following siNrf2 transfection into endothelial cells, the anti­inflammatory and antioxidant stress effects of LBP were decreased. LBP was found to enhance the expression of the Keap1/Nrf2 antioxidant stress signaling pathway in endothelial cells, decreasing oxidative stress and the inflammatory response. Moreover, LBP improved the antioxidant stress ability of endothelial cells and alleviated injury of myocardial vascular tissue, thereby protecting the cardiovascular system.

The predictive value of the ratio of the product of neutrophils and hemoglobin to lymphocytes in non-muscular invasive bladder cancer patients with postoperative recurrence.

PURPOSE: The purpose of this study was to explore the predictive value of the ratio of the product of neutrophils and hemoglobin to lymphocytes (NHL) in patients with non-muscular invasive bladder cancer (NMIBC). MATERIALS AND METHODS: We retrospectively collected clinical and pathological data of patients with NMIBC who underwent transurethral resection of bladder tumor (TURBT) at our hospital between 2013 and 2018. The ratio of neutrophils to lymphocytes (NLR), the Systemic Immune Inflammation Index (SII), and NHL were obtained based on routine blood settlement within a week before surgery. The receiver operating characteristic curve was used to determine the optimal cutoff value of each index, and different groups were grouped accordingly. Kaplan-Meier survival curve and Cox regression model were used to study the factors affecting the prognosis of NMIBC patients. RESULTS: There was significant difference in recurrence-free survival (RFS) rate between the high NLR group and the low NLR group, the high SII group and the low SII group, and the high NHL group and the low NHL group. Cox univariate regression analysis showed that tumor number, tumor size, tumor pathological grade, tumor pathological stage, NLR, SII, and NHL were related to postoperative RFS in patients with NMIBC. The tumor number, tumor pathological grade, SII, and NHL were independent predictors of RFS in multivariate analysis. CONCLUSIONS: The preoperative clinical inflammatory indexes NLR, SII, and NHL have certain predictive value for postoperative RFS in NMIBC patients.

[Betaine induces apoptosis of C4-2B prostate cancer cells via inhibiting PI3K/AKT/NF-κB signaling pathway].

Objective To investigate the inhibitory effect of betaine (BET) on the proliferation of C4-2B prostate cancer cells and its possible mechanism. Methods C4-2B cells were treated with 0, 100, 200, 400, 600, 800 mmol/L BET. CCK-8 assay was used to assess the cell proliferation, plate cloning formation assay to detect clone formation ability and flow cytometry to evaluate cell apoptosis, and the cell morphological alteration was observed by microscopy. The protein expression of BAX, Bcl2, cleaved caspase 3 (c-caspase-3), phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), and NF-κB p65 were detected by Western blotting, and the changes of BAX, Bcl2, c-caspase-3, and NF-κB p65 proteins were further verified after the cells were treated with NF-κB pathway inhibitor BAY11-7082. Results BET inhibited the proliferation of C4-2B cells in a dose-dependent manner. The 50% inhibitory concentration (IC50) was 422.7 mmol/L after the cells were treated with BET for 48 hours. Compared with the control group (0 mmol/L BET treatment), the proliferation of C4-2B cells was inhibited along with morphological changes, decreased clone formation ability and increased apoptosis rate in 200, 300, 400 mmol/L BET treated groups. Meanwhile, the protein expression of BAX and c-caspase-3 were up-regulated and Bcl2, PI3K, AKT and NF-κB p65 were down-regulated in 300, 400 mmol/L BET groups as compared with the control group. After BAY11-7082 treatment alone, Bcl2, BAX, c-caspase-3, NF-κB p65 protein expression trend was consistent with that of the 300 mmol/L BET treated group, and Bcl2, NF-κB p65 protein expression levels were lower and BAX and c-caspase-3 protein expression levels were higher in BET combined with BAY11-7082 treated group. Conclusion BET can inhibit C4-2B cell proliferation and induce its apoptosis by blocking PI3K/AKT/NF-κB signaling pathway.

The meta-analysis of the effect of 68Ga-PSMA-PET/CT diagnosis of prostatic cancer compared with bone scan.

BACKGROUND: 68Ga-PSMA-PET/CT (positron emission tomography/computed tomography) is a promising method for prostate cancer (PC) detection. However, the ability of 68Ga-PSMA-PET/CT to detect malignant bone lesions, and whether this method is superior to the existing bone imaging methods are still lack of systematic analysis. PURPOSE: To evaluate the value of 68Ga-PSMA-PET/CT and bone scan in clinical diagnosis of prostatic cancer from the perspective of evidence-based medicine. METHODS: PubMed, The Cochrane Library, EMBASE, Springer Link, Sinomed, CNKI, Wanfang database, and CQVIP database were searched to find the satisfactory studies that needed systematic review of trials and compared the value of 68Ga-PSMA-PET/CT and bone scan. All studies published from inception to March 31, 2020. According to the inclusion and exclusion criteria, 2 reviewers independently evaluated and extracted the literature. Review Manager 5.3 was applied to evaluate the included literature quality. The heterogeneity of the included literature was tested by Meta Disc 1.4, and the effect model was selected according to the heterogeneity test results, and the sensitivity (SEN), specificity (SPE), PLR, NLR and diagnostic odds ratio (DOR) were analyzed. After testing the heterogeneity results of literature by using the 95% confidence interval and the forest map. RESULTS: A total of 4 studies were eligible for inclusion in the meta-analysis, which included 318 patients, 120 cases with bone metastasis and 198 cases without bone metastasis. The results of summary evaluation for 68Ga-PSMA-PET/CT and bone scan in diagnosis of prostatic cancer as follow respectively: The SEN were 0.97 and 0.86; the SPE were 1.00 and 0.87; the DOR were 1468.33 and 36.23; PLR were 88.45 and 6.67; NLR were 0.05 and 0.19; and the area under curve (AUC) and 95% CI were 0.9973 (1.0000-0.9927) and 0.8838 (0.9584-0.8092). CONCLUSION: By comparing the diagnostic results of 68Ga-PSMA-PET/CT and bone scan imaging diagnosis methods, the 68Ga-PSMA-PET/CT has a higher SEN and SPE than bone scan, and it has a higher diagnostic efficiency for prostate cancer bone metastasis, which is worthy of clinical application.

Effects of norepinephrine on colonic tight junction protein expression during heat stress.

Stress induced by changes in the internal or external environment in humans and animals leads to intestinal epithelial damage, in a manner that is associated with impaired intestinal barrier function. However, the role of the stress hormone norepinephrine (NE) in impairments in barrier function remains poorly understood. In the present study, a rat heat-exposed model was used to observe changes in the tight junction proteins Occludin and zonula occludens-1 (ZO-1), in addition to those in protease-activated receptor 2 (PAR-2) and transient receptor potential ankyrin 1 channel (TRPA1) in colon. The levels of plasma NE were detected using an ELISA kit. Different concentrations of NE were used to culture the human colon cell line Caco-2 for 6 and 24 h to investigate the cell viability using Cell Counting Kit-8 assay, whilst the expression levels of Occludin, ZO-1, PAR-2 and TRPA1 were examined using western blotting and immunofluorescence in Caco-2 cells and immunohistrochemistry in rat colon tissues. Although there was no clear histological damage to the rat colonic mucosa, there were decreased expression levels of tight junction proteins Occludin and ZO-1 after heat exposure. In addition, PAR-2 expression was increased by heat exposure. It was found that TRPA1 expression was concentrated to the luminal surface of the colon in the heat exposed group compared with that in the control group. After the administration of increasing concentrations of NE for 6 h, treatment did not affect cell viability. Furthermore, after application of NE for 24 h, cell viability gradually increased as the NE concentration was elevated from 10 to 100 µM. However, no significant increase in viability was observed when the cells were treated with 120 and 160 µM NE. Occludin expression was decreased when 10 µM NE was applied for 6 or 24 h. By contrast, 60 µM NE significantly downregulated Occludin expression in the 6 h group, but caused an insignificant decrease in the 24 h group. It was found that ZO-1 expression was upregulated after treatment with 10 µM NE for 6 h, whilst downregulation was observed after treatment with 10 µM NE for 24 h. PAR-2 protein expression was increased after application of NE for both 6 and 24 h, but not after treatment with 60 µM NE. In addition, TRPA1 expression was not affected by the treatment of NE, but increased positive staining was observed on the luminal side of the mucosa, which appeared to be concentrated in the cells of the luminal side in the rat colon after heat exposure. Collectively, the present results suggested that expression of tight junction proteins Occludin and ZO-1, in addition to that of PAR-2, can be regulated by NE, which may contribute to impairments in barrier function observed during heat stress.

LncRNA XIST modulates 5-hydroxytrytophan-induced visceral hypersensitivity by epigenetic silencing of the SERT gene in mice with diarrhea-predominant IBS.

Diarrhea-predominant irritable bowel syndrome (IBS-D) is a prevalent gastrointestinal disorder with a high incidence in children. The role of long non-coding RNAs (lncRNAs) in gastrointestinal diseases has been previously highlighted. Nevertheless, the underlying regulatory mechanism of lncRNA X inactivate-specific transcript (XIST) in IBS-D requires further studies. Thus, the present study was conducted with the main objective of elucidating the underlying mechanism of lncRNA XIST in visceral hypersensitivity in IBS-D. An in vivo mouse model of IBS-D was constructed via rectal perfusion of acetic acid. Next, in order to evaluate the effect of lncRNA XIST on the development of visceral hypersensitivity in IBS-D, different vector plasmids were injected into mice along with rectal mucosal epithelial cells, followed by the measurement of abdominal withdrawal reflex (AWR) score, counts of peristaltic wave, abdominal wall contraction and defecation particles. Furthermore, luciferase reporter assay, FISH, RIP and ChIP assays were conducted to determine the interactions between lncRNA XIST and SERT. Subsequently, MS-PCR was adopted to test the methylation level of SERT promoter. 5-hydroxytrytophan (HT) content in rectal tissues was detected using immunohistochemistry. The IBS-D mouse models presented with a high expression of lncRNA XIST along with low expression of SERT. LncRNA XIST was observed to recruit methylase DNMT1, DNMT3A and DNMT3B to promote SERT promoter methylation, reducing its expression. Restoration of lncRNA XIST resulted in increased AWR score, counts of peristaltic wave, abdominal wall contraction and defecation particles along with stimulated 5-HT expression and SERT methylation level, while downregulation of lncRNA XIST reversed these effects. In conclusion, the key findings from our study indicated that lncRNA XIST acts as a regulator in 5-HT-induced visceral hypersensitivity in mice with IBS-D, providing a new insight into the regulatory effect of lncRNA XIST and its epigenetic diagnostic and therapeutic properties in IBS-D.

Homocysteine accelerates atherosclerosis by inhibiting scavenger receptor class B member1 via DNMT3b/SP1 pathway.

Homocysteine (Hcy) is an independent risk factor for atherosclerosis, which is characterized by lipid accumulation in the atherosclerotic plaque. Increasing evidence supports that as the main receptor of high-density lipoprotein, scavenger receptor class B member 1 (SCARB1) is protective against atherosclerosis. However, the underlying mechanism regarding it in Hcy-mediated atherosclerosis remains unclear. Here, we found the remarkable inhibition of SCARB1 expression in atherosclerotic plaque and Hcy-treated foam cells, whereas overexpression of SCARB1 can suppress lipid accumulation in foam cells following Hcy treatment. Analysis of SCARB1 promoter showed that no significant change of methylation level was observed both in vivo and in vitro under Hcy treatment. Moreover, it was found that the negative regulation of DNMT3b on SCARB1 was due to the decreased recruitment of SP1 to SCARB1 promoter. Thus, we concluded that inhibition of SCARB1 expression induced by DNMT3b at least partly accelerated Hcy-mediated atherosclerosis through promoting lipid accumulation in foam cells, which was attributed to the decreased binding of SP1 to SCARB1 promoter. In our point, these findings will provide novel insight into an epigenetic mechanism for atherosclerosis.

Protective functions of Lycium barbarum polysaccharides in H2O2-injured vascular endothelial cells through anti-oxidation and anti-apoptosis effects.

Cell injury in the cardiovascular endothelia caused by oxidative stress is among the major inducers of endothelium dysfunction and serves an important role in initiating cardiovascular diseases (CVDs). Therefore, protecting and improving the normal function of endothelial cells are considered key measures against CVDs. As a traditional Chinese medicinal component, Lycium barbarum is regarded to have high medicinal value. The present study aimed to investigate the potential anti-apoptosis and anti-oxidation effects of Lycium barbarum polysaccharides (LBPs) on injured rat artery endothelial cells, to demonstrate the experimental and medicinal values of LBPs. In the present study, the aortic endothelial cells of rats were cultivated and randomly divided into five groups: A control group, H2O2-injured group (H2O2 group), H2O2+LBPs (110 µg/ml) group (low-dose group, LT), H2O2+LBPs (220 µg/ml) group (medium-dose group, MT) and H2O2+LBPs (440 µg/ml) group (high-dose group, HT). Among these, the activity of superoxide dismutase (SOD), and the levels of malondialdehyde (MDA) and nitric oxide (NO) were detected by colorimetry. Additionally, the expression of B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) were detected by western blotting. It was observed that SOD activity and NO content decreased while MDA content increased significantly in the H2O2 group (P<0.05 vs. control); that SOD activity in the MT and HT group, and NO content in all three LBP groups were increased, while MDA content in the three LBP groups was decreased, compared with the H2O2 group (all P<0.05); that Bcl-2 expression decreased significantly in the H2O2 group while the expression of Bax increased significantly compared with the control group (both P<0.05); and that Bcl-2 expression in all three LBP groups increased, while Bax expression in the MT and HT groups decreased compared with the H2O2 group (all P<0.05), with these altered Bax levels being statistically similar to those in the control group (P>0.05). On light microscopy, the cells in the control group exhibited spindle-shaped morphology, consistent sizes, defined boundaries, and distinct nuclei of equivalent sizes with round or oval morphology. Additionally, the chromatin in the nuclei was evenly distributed, and all cells were adhered in a paving-stone arrangement. Notably, only few cells died. Conversely, the cells in the H2O2 group exhibited signs of damage and enlarged gaps, and focal cells died. In the HT group, the cells once again appeared adherent and exhibited similar morphological status to the normal cells. Overall, these results indicate that LBPs serve a protective role in oxidative-injured vascular endothelial cells through anti-apoptosis and anti-oxidation effects.

Role of EGFR/ErbB2 and PI3K/AKT/e-NOS in Lycium barbarum polysaccharides Ameliorating Endothelial Dysfunction Induced by Oxidative Stress.

Lycium barbarum polysaccharides (LBP) are the major ingredients of wolfberry. In this study, we investigated the role of LBP in endothelial dysfunction induced by oxidative stress and the underlying mechanisms using thoracic aortic endothelial cells of rat (RAECs) as a model. We found that Ang II inhibits cell viability of RAECs with 10-6mol/L of Ang II treatment for 24h most potential (P<0.05), the level of reactive oxygen species (ROS) is increased by Ang II treatment (P<0.01), and the expression of Occludin and Zonula occludens-1 (ZO-1) is decreased by Ang II treatment (P<0.05). However, preincubation of cells with LBP could inhibit the changes caused by Ang II, LBP increased cell viability (P<0.05), decreased the level of ROS (P<0.01), and up-regulated the expression of Occludin (P<0.05) and ZO-1. In addition, Ang II treatment increased the expression of EGFR and p-EGFR (Try1172) and which can be inhibited by LBP. On the contrary, expression of ErbB2, p-ErbB2 (Try1248), PI3K, p-e-NOS (Ser1177) (P<0.05), and p-AKT (Ser473) (P<0.05) was inhibited by Ang II treatment and which can be increased by LBP. Treatment of the cells with inhibitors showed that the regulation of p-e-NOS and p-AKT expression by Ang II and LBP can be blocked by PI3K inhibitor wortmannin but not EGFR and ErbB2 inhibitor AC480. Taken together, our results suggested that LBP plays a critical role in maintaining the integrality of blood vessel endothelium through reduced production of ROS via regulating the activity of EGFR, ErbB2, PI3K/AKT/e-NOS, and which may offer a novel therapeutic option in the management of endothelial dysfunction.