Circular RNA circCCDC85A inhibits breast cancer progression via acting as a miR-550a-5p sponge to enhance MOB1A expression.

BACKGROUND: A growing body of evidence indicates that abnormal expression of circular RNAs (circRNAs) plays a crucial role by acting as molecular sponges of microRNAs (miRNAs) in various diseases, including cancer. In this study, we explored whether circCCDC85A could function as a miR-550a-5p sponge and influence breast cancer progression. METHODS: We detected the expression of circCCDC85A in breast cancer tissues and cells using fluorescence in situ hybridization (FISH) and quantitative reverse transcription polymerase chain reaction (qRT-PCR). CCK-8 and colony formation assay were used to detect the proliferative ability of breast cancer cells. Wound healing assay and transwell migration and invasion assays were used to detect the migrative and invasive abilities of breast cancer cells. We also examined the interactions between circCCDC85A and miR-550a-5p using FISH, RNA-binding protein immunoprecipitation (RIP), and luciferase reporter assay. Moreover, we performed luciferase reporter assay, qRT-PCR, and Western blot to confirm the direct targeting of miR-550a-5p to MOB1A. RESULTS: The expression of circCCDC85A in breast cancer tissues was obviously lower than that in normal breast tissues. Over-expression of circCCDC85A substantially inhibited the proliferative, migrative, and invasive ability of breast cancer cells, while knocking down of circCCDC85A enhanced the aforementioned properties of breast cancer cells. Moreover, enforced expression of circCCDC85A inhibits the oncogenic activity of miR-550a-5p and increases the expression of MOB1A targeted by miR-550a-5p. Further molecular mechanism research showed that circCCDC85A may act as a molecular sponge for miR-550a-5p, thus restoring miR-550a-5p-mediated targeting repression of tumor suppressor MOB1A in breast cancer cells. CONCLUSION: Our findings provide novel evidence that circCCDC85A inhibits the progression of breast cancer by functioning as a molecular sponge of miR-550a-5p to enhance MOB1A expression.

Magnetic polyimide nanocomposite for analysis of parabens in cooking wine by magnetic solid-phase extraction coupled with gas chromatography - Mass spectrometry.

A magnetic polyimide (PI) nanocomposite has been synthesized by phase inversion of PI and simultaneous encapsulation of Fe3O4 nanoparticles. The Fe3O4/PI nanocomposite was characterized by a variety of characterization techniques, including infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, nitrogen adsorption-desorption isotherms, and vibrating sample magnetometry. The results showed that the prepared nanocomposite had a homogeneous structure, adequate specific surface area (76.1 m2/g) and high saturation magnetization (42.9 emu/g). Using parabens as model analytes, the performance of the Fe3O4/PI nanocomposite as an adsorbent for magnetic solid-phase extraction (MSPE) was evaluated. The extracted parabens were desorbed and determined by gas chromatography-mass spectrometry (GC-MS). The parameters affecting the extraction and desorption efficiency of parabens were optimized. Under the optimal conditions, the developed MSPE/GC-MS method was successfully applied to the determination of parabens in cooking wine. The MSPE/GC-MS method exhibited broad linearity (0.2-100 µg/L), low detection limits (0.04-0.05 µg/L), and satisfactory extraction recoveries (79.2 %-113.3 %) with relative standard deviations (RSDs) ranging from 0.7 % to 10.4 %. For real cooking wine samples, the spiked recoveries ranged from 91.7 % to 118.7 % with RSDs of 1.0 %-11.2 %. The results demonstrated that the Fe3O4/PI nanocomposite was an effective adsorbent, and this work provides a novel reference for the easy preparation of magnetic adsorbent materials.

Optimization of Ultrahigh-Throughput Screening Assay for Protein Engineering of d-Allulose 3-Epimerase.

d-Allulose is the corresponding epimer of d-fructose at the C-3 position, which exhibits a similar taste and sweetness to sucrose. As a low-calorie sweetener, d-allulose has broad application prospects in the fields of medicine, food, and so on. Currently, the production method of d-allulose is mainly the enzymatic conversion of d-fructose by d-allulose 3-epimerase (DAEase). However, the limited specific activity and thermal stability of DAEase restrict its industrial application. Herein, an ultrahigh-throughput screening assay based on the transcription factor PsiR was extensively optimized from the aspects of culture medium components, screening plasmid, and expression host, which enhanced the correction between the fluorescent readout and the enzyme activity. Then, the error-prone PCR (epPCR) library of Clostridium cellulolyticum H10 DAEase (CcDAEase) was screened through the above optimized method, and the variant I228V with improved specific activity and thermal stability was obtained. Moreover, after combining two beneficial substitutions, D281G and C289R, which were previously obtained by this optimized assay, the specific activity of the triple-mutation variant I228V/D281G/C289R reached up to 1.42-fold of the wild type (WT), while its half-life (T1/2) at 60 °C was prolonged by 62.97-fold. The results confirmed the feasibility of the optimized screening assay as a powerful tool for the directed evolution of DAEase.

Ghrelin in the lateral parabrachial nucleus influences the excitability of glucosensing neurons, increases food intake and body weight.

Ghrelin plays a pivotal role in the regulation of food intake, body weight and energy metabolism. However, these effects of ghrelin in the lateral parabrachial nucleus (LPBN) are unexplored. C57BL/6J mice and GHSR-/- mice were implanted with cannula above the right LPBN and ghrelin was microinjected via the cannula to investigate effect of ghrelin in the LPBN. In vivo electrophysiological technique was used to record LPBN glucose-sensitive neurons to explore potential udnderlying mechanisms. Microinjection of ghrelin in LPBN significantly increased food intake in the first 3 h, while such effect was blocked by [D-Lys3]-GHRP-6 and abolished in GHSR-/- mice. LPBN ghrelin microinjection also significantly increased the firing rate of glucose-excited (GE) neurons and decreased the firing rate of glucose-inhibited (GI) neurons. Additionally, LPBN ghrelin microinjection also significantly increased c-fos expression. Chronic ghrelin administration in the LPBN resulted in significantly increased body weight gain. Meanwhile, no significant changes were observed in both mRNA and protein expression levels of UCP-1 in BAT. These results demonstrated that microinjection of ghrelin in LPBN could increase food intake through the interaction with growth hormone secretagogue receptor (GHSR) in C57BL/6J mice, and its chronic administration could also increase body weight gain. These effects might be associated with altered firing rate in the GE and GI neurons.

Up-regulated circular RNA VANGL1 contributes to progression of non-small cell lung cancer through inhibition of miR-195 and activation of Bcl-2.

Circular RNAs (circRNAs), a group of non-coding RNAs, play an important role in cancer biology, and in the present study, we aimed to clarify the expression profiles and biological functions of circRNA circVANGL1 in non-small cell lung cancer (NSCLC). The results showed that circVANGL1 was overexpressed in human NSCLC tissues and cell lines. circVANGL1 expression was closely associated with tumor size, TNM stage and overall survival of NSCLC patients. Further loss-of-function analysis revealed that knockdown of circVANGL1 inhibited proliferation and induced apoptosis in NSCLC cell lines. The migration and invasion of NSCLC cells were also suppressed by circVANGL1 knockdown. In addition, we predicted that circVANGL1 might serve as a competing endogenous RNA (ceRNA), becoming a sink for miR-195, thereby modulating the expression of Bcl-2 in NSCLC cells. Rescue experiments demonstrated that miR-195 inhibitor abrogated the beneficial role of circVANGL1 knockdown in NSCLC cells. Taken together, we conclude that circVANGL1 functions as an oncogene to promote NSCLC progression partly through miR-195/Bcl-2 axis, which might be a novel therapeutic target for NSCLC patients.

L-carnitine ameliorates peripheral neuropathy in diabetic mice with a corresponding increase in insulin­like growth factor­1 level.

Diabetic peripheral neuropathy (DPN) is one of the common complications in diabetes, affecting more than half of patients with diabetes. L­carnitine (LC) was recently demonstrated to serve a positive role in ameliorating DPN. Therefore, the aim of the present study was to investigate the underlying mechanisms of LC in ameliorating DPN. Male Kunming mice were randomly assigned into five groups, including the control group, diabetes mellitus group, pre­treatment group, treatment group and post­treatment group. Type 2 diabetes was induced in mice using a combination of high­fat diet and streptozotocin injection. Subsequently, peripheral neuropathy was measured and the levels of LC, insulin and insulin­like growth factor­1 (IGF­1) were detected. When diabetic mice were treated with LC, the levels of IGF­1 in the plasma and pancreas were increased. In addition, hyperalgesia, as determined by the tail­flick test as well as food intake, body weight and blood glucose levels were decreased. An amelioration of demyelination, axonal atrophy and mitochondria swelling in the nerve fibres of diabetic mice was also observed. The present study demonstrated that LC ameliorated peripheral neuropathy in type 2 diabetic mice and the effect of LC may in part be mediated by an increase in local and circulatory IGF­1 levels.

Recombinant betatrophin (Angptl­8/lipasin) ameliorates streptozotocin­induced hyperglycemia and ß­cell destruction in neonatal rats.

Betatrophin [also known as lipasin, angiopoietin­like 8 (ANGPTL8), refeeding induced in fat and liver (RIFL), or hepatocellular carcinoma­associated gene TD26], a 22­kDa protein in the angiopoietin­like family, is a liver­derived hormone that promotes pancreatic ß­cell proliferation and lipid metabolism. The aim of the present study was to investigate the effect of recombinant betatrophin on ß­cell regeneration in a neonatal streptozotocin (STZ)­induced diabetic rat model. One­day­old Wistar rats were injected with STZ (100 mg/kg), followed by intraperitoneal administration of betatrophin to the STZ­injected rats for 6 days. Plasma glucose and body weight were monitored. On days 4 and 7, expression levels of pancreatic duodenal homeobox gene­1 (PDX­1), the Bax/B­cell lymphoma­2 (Bcl­2) ratio and plasma insulin were assessed, and the ß­cell proliferation rate was determined. Pancreatic islet area and number were determined at 10 weeks. It was found that betatrophin treatment alleviated STZ­induced hyperglycemia, elevated pancreatic expression levels of Bcl­2, PDX­1, plasma insulin levels and the ß­cell proliferation rate on days 4 and 7. Long­term betatrophin treatment improved glucose tolerance, associated with improved plasma insulin levels and ß­cell mass. These results suggest that early administration of betatrophin promotes ß­cell proliferation in STZ­induced diabetic neonates and prevents the development of diabetes in adults.

Effects of Lifestyle Modification and Anti-diabetic Medicine on Prediabetes Progress: A Systematic Review and Meta-Analysis.

Background: Pre-diabetes is a risk factor for full-blown diabetes; it presents opportunities to prevent the actual diseases. It is therefore essential to identify effective preventive strategies, and to clarify the direction of future research. Methods: PubMed, Embase and Cochrane Central Register of Controlled Trials were searched using key terms (Supplementary Table 1). We applied network meta-analysis to multiple comparisons among various diabetic preventive strategies, including lifestyle and pharmacological interventions; traditional meta-analysis for the synthesis of basal metabolic changes after interventions; and trial sequential analysis for determinations as to whether analysis conclusions meet expectations. Results: We included 32 randomized controlled trials comprising 43,669 patients and 14 interventions in the meta-analysis. Both lifestyle modifications and anti-diabetic medications improved physical conditions, including weight loss, blood glucose, and blood pressure. Network meta-analysis suggested that the progression of diabetes could be delayed to varying degrees by lifestyle and pharmacological interventions, except for angiotensin-converting enzyme inhibitors, statins, sulfonylureas and vitamin D. The risk ratios (RR) [95% credible interval (CrI)] compared with control were: GLP-1RAs 0.28 (0.15, 0.50), Orlistat 0.33 (0.18, 0.55), TZM 0.33 (0.16, 0.63), TZD 0.39 (0.27, 0.53), LST 0.54 (0.32, 0.88), lifestyle 0.58 (0.49, 0.67), LSM 0.62 (0.45, 0.80), GI 0.66 (0.46, 0.88), SU 0.67 (0.40, 1.00), Vitamin D 0.91 (0.59, 1.40), ACEI 0.93 (0.62, 1.40), statins 1.20 (0.84, 1.60). Conclusions: In adults with pre-diabetes, firm evidence supports the notion that lifestyle modifications and metformin reduces the incidence of diabetes with an average of 20% relative risk reduction, while statins increase the relative risk 20%. We found that lifestyle modifications, promising long-term strategies involving three factors (nutrition, exercise, and weight loss) contribute to health by reducing BMI, body weight, waist and hip circumference, systolic and diastolic pressure, fasting, and 2-h postprandial blood glucose, total cholesterol and by increasing HDL. We made this determination using TSA, avoiding further waste of experimental resources.

The burden of air pollution and weather condition on daily respiratory deaths among older adults in China, Jinan from 2011 to 2017.

The health effects of short-term exposure to air pollutants on respiratory deaths and its modifiers such as meteorological indexes have been widely investigated. However, most of the previous studies are limited to single pollutants or total respiratory deaths, and their findings are inconsistent.To comprehensively examine the short-term effects of air pollutants on daily respiratory mortality.Our analysis included 16,931 nonaccidental respiratory deaths (except lung cancer and tuberculosis) among older adults (>65 years) from 2011 to 2017 in Jinan, China. We used a generalized additive Poisson models adjusted for meteorology and population dynamics to examine the associations between air pollutants (particulate matter with an aerodynamic diameter of b2.5µm [PM2.5], particulate matter with an aerodynamic diameter of b10µm [PM10], SO2, NO2, O3) and daily mortality for the total patients, males, females, chronic airway diseases, pneumonia patients, and rest patients in Jinan.Outdoor air pollution was significantly related to mortality from all respiratory diseases especially from chronic airway disease in Jinan, China. The effects of air pollutants had lag effects and harvesting effects, and the effects estimates usually reached a peak at lag 1 or 2 day. An increase of 10 µg/m or 10 ppb of PM2.5, PM10, SO2, NO2, and O3 corresponds to increments in mortality caused by chronic airway disease of 0.243% (95% confidence interval [CI]: -0.172-0.659) at lag 1 day, 0.127% (95% CI: -0.161-0.415) at lag 1 day, 0.603% (95% CI: 0.069-1.139) at lag 3 day, 0.649% (95% CI: -0.808-2.128) at lag 0 day and 0.944% (95% CI: 0.156-0.1598) at lag 1 day, respectively. The effects of air pollutants were usually greater in females and varied by respiratory subgroups. Spearman correlation analysis suggested that there was a significant association between meteorological indexes and air pollutants.Sex, age, temperature, humidity, pressure, and wind speed may modify the short-term effects of outdoor air pollution on mortality in Jinan. Compared with the other pollutants, O3 had a stronger effect on respiratory deaths among the elderly. Moreover, chronic airway diseases were more susceptible to air pollution. Our findings provided new evidence for new local environmental and health policies making.

Correction to: Systematic Review and Meta-Analysis of the Change in Ghrelin Levels After Roux-en-Y Gastric Bypass.

This article was initially published with incorrect copyright information. Upon publication of this correction, the copyright of this article changed to "The Author(s)." The original article has been corrected.

Systematic Review and Meta-analysis of the Change in Ghrelin Levels After Roux-en-Y Gastric Bypass.

BACKGROUND: Roux-en-Y gastric bypass (RYGB) is considered effective for weight loss and for treatment of many obesity-related metabolic diseases. Ghrelin is an essential orexigenic peptide that plays an indispensable role in controlling body weight and energy homeostasis of post-operative patients. This systematic review and meta-analysis aimed to investigate changes in the level of fasting total ghrelin following RYGB. METHODS: A systematic literature search of PubMed, EMBASE, and the Cochrane Library until April 2018 with keywords "ghrelin" and "gastric bypass" was performed in accordance with the MOOSE guidelines and PRISMA statement. Three reviewers independently selected the studies and extracted data. Quality assessment of the included studies was undergone. A random effects model was employed to calculate overall effect sizes. Subgroup analyses and meta-regression were subsequently performed. RESULTS: Sixteen studies with 325 patients were included. We found ghrelin levels had an increasing tendency (SMD = 0.30; 95% CI = 0.04 to 0.57) despite moderate heterogeneity (I2 = 58%). Subsequent subgroup analysis indicated that ghrelin levels decreased (SMD = - 0.49; 95% CI = - 0.98 to 0.00) in the short term (≤ 3 months) and increased (SMD = 0.46; 95% CI = 0.22 to 0.69) in the long term (> 3 months) after RYGB. Meta-regression showed that gastric pouch volume, alimentary limb length and biliopancreatic limb length were not associated with changes in ghrelin levels. CONCLUSION: Fasting total ghrelin levels decreased in the short term (≤ 3 months) and increased in the long term (> 3 months) after RYGB.

Proliferating cell nuclear antigen promotes cell proliferation and tumorigenesis by up-regulating STAT3 in non-small cell lung cancer.

Proliferating cell nuclear antigen (PCNA) functions as a bridging molecule, which targets proteins that have distinct roles in cell growth. The expression of PCNA is dysregulated in some tumors and takes part in the progression of oncogenesis. However, the roles of PCNA in the progression of non-small cell lung cancer (NSCLC) remain unknown. The present study aimed to investigate the function of PCNA in the occurrence and development of NSCLC and its underlying molecular mechanisms. Western blotting, RT-PCR, and immunohistochemistry assays were used to detect the expression pattern of PCNA in NSCLC tissues and cells. A log rank test was performed to compare the overall survival (OS) of patients with high/low expression of PCNA. Besides, the relationship between PCNA and signal transducer and activator of transcription-3 (STAT3) proteins were evaluated. Then, MTT, flow cytometry, clonal formation, and in vivo xenograft assays were conducted to investigate the effects of PCNA/STAT3 on cell growth, clonal formation, apoptosis, and tumorigenesis. Results showed that PCNA expression was elevated in NSCLC tissues and cells and it could combine with STAT3 and increased its expression and phosphorylation. Moreover, the expression of PCNA showed a positive correlation with the TNM grade and occurrence rate of the lymphatic metastasis and poor prognosis of NSCLC patients. Overexpression of PCNA promoted cell proliferation, clonal formation, and tumorigenesis in lung cancer cells and inhibited cell apoptosis. In contrast, these effects were inhibited when knockdown of STAT3. In conclusion, this study demonstrates that PCNA functions as an oncogene in the progression of NSCLC through up-regulation of STAT3. These findings point to a potentially new therapeutic strategy for NSCLC.

ML-7 attenuates airway inflammation and remodeling via inhibiting the secretion of Th2 cytokines in mice model of asthma.

Previous studies have indicated that smooth muscle myosin light chain kinase (MLCK) has a prominent role in the regulation of smooth muscle contraction, which tends to be upregulated in asthma. In recent years, numerous studies have reported that MLCK is intimately connected with the immunoregulatory mechanism of T cells. The imbalance of T helper type 1 cells (Th1)/Th2 constitutes the immune­associated pathological basis of chronic asthma. Th2­associated cytokines, including interleukin­4, ­5, ­13, ­25 and ­33, are involved in airway inflammation, hyperresponsiveness and remodeling, which leads to a progressive decline in lung function. The purpose of the present study was to verify whether inhibition of bronchial MLCK attenuated the expression Th2­associated cytokines in asthmatic mice, including the above­mentioned ones. Female BALB/c mice were used to establish an ovalbumin (OVA)­induced model of asthma, of which one group was treated with the MLCK inhibitor (5-iodonaphthalene-1-sulfonyl) homopiperazine (ML­7). The inhibitor of MLCK, ML­7 attenuated airway inflammation and remodeling by reducing inflammatory cell infiltration and the secretion of Th2 cytokines in mice model of asthma, which may represent a promising therapeutic strategy for asthma.

Neuropeptide Y-Positive Neurons in the Dorsomedial Hypothalamus Are Involved in the Anorexic Effect of Angptl8.

Angiopoietin-like protein 8 (Angptl8), a recently identified member of the angiopoietin-like protein family (ANGPTLs), is a 22-kDa peptide synthesized in the liver. It participates in lipid metabolism by inhibiting lipoprotein lipase (LPL) activity, consequently increasing the triglyceride levels. Despite evidence that Angptl8 is involved in feeding control, the underlying mechanisms are unclear. Central and peripheral injections of Angptl8 significantly decreased food intake. Angptl8 was widely expressed in appetite-related nuclei, including the paraventricular nucleus (PVN), the dorsomedial hypothalamus (DMH), the ventromedial hypothalamus, and the arcuate nucleus (ARC) in the hypothalamus. Peripheral Angptl8 administration decreased c-Fos-positive neurons in the DMH. Central Angptl8 administration decreased c-Fos-positive neurons in the DMH and PVN but increased these neurons in the ARC. Angptl8 inhibited appetite via neuropeptide Y (NPY) neurons in the DMH. Furthermore, the chronic administration of Angptl8 decreased body weight gain and altered adipose tissue deposits. Nevertheless, neither peripheral nor central Angptl8 influenced the brown adipose tissue (BAT) morphology or uncoupling protein 1 (Ucp-1) expression in BAT. Taken together, these data suggested that Angptl8 modulates appetite and energy homeostasis.

The Correlation Between Circulating Ghrelin and Insulin Resistance in Obesity: A Meta-Analysis.

Background: Ghrelin, a peptide mainly produced by stomach X-A cells. It plays a pivotal role in the regulation of food intake and energy metabolism, including glucose metabolism and insulin sensitivity. However, the correlation between circulating ghrelin levels and insulin resistance in obesity remained uncertain. This meta-analysis aimed to clarify the association between ghrelin and IR in obesity. Methods: A systematic literature search was performed using PubMed, EMBASE, Cochrane Library and Web of Science until April 18, 2018 with the keywords "ghrelin" and "insulin resistance." Two independent reviewers selected studies and assessed data. Subgroup analyses were performed to search for sources of heterogeneity. Funnel plots and Egger's test were used to detect publication bias. A random-effects model was used to calculate the pooled effect size. Results: Ten studies with 546 participants were included in this meta-analysis. We found that ghrelin levels were negatively correlated with IR in obese individuals. (r = -0.31; 95% CI: -0.45 to -0.18). Subgroup analysis revealed that circulating ghrelin levels were significantly negatively correlated with IR in people with normal fasting blood glucose (FBG) (<6.9 mmol/dl) (r = -0.28; 95% CI: -0.47 to -0.09, I 2 = 39.5%), while there was no relationship between circulating ghrelin levels and IR in the high FBG group (>6.9 mmol/dl) (r = -0.15; 95% CI: -0.33 to 0.03, I 2 = 0.0%). Publication bias was insignificant (Egger's test: P = 0.425). Conclusion: In obesity, circulating ghrelin levels were significantly negative correlated with insulin resistance in individuals with normal fasting blood glucose.

Ghrelin Receptor Is Required for the Effect of Nesfatin-1 on Glucose Metabolism.

Studies of nesfatin-1 in glucose metabolism have become a topic of interest recently, however, the specific receptor for nesfatin-1 has not yet been identified. Some studies hinted at a connection between nesfatin-1 and the ghrelin receptor, growth hormone secretagogue receptor. Therefore, we aimed to study the role of GHSR in the glycemic effects of nesfatin-1 as well as its downstream pathways. We employed C57/BL6 mice (wild type and GHSR knockout mice) eating a normal chow diet and a high fat diet in this study, and the experimental technique included western blot, real-time PCR, immunofluorescence and ELISA. We found that in mice fed a normal chow diet (NCD), nesfatin-1 improved glucose tolerance, up-regulated AKT kinase (AKT) mRNA levels and phosphorylation and GLUT4 membrane translocation in skeletal muscle. These effects were blocked by co-injection of GHSR antagonist [D-Lys3]-GHRP-6 and were attenuated in GHSR knockout mice. In mice fed high-fat diet (HFD), nesfatin-1 not only exerted the effects observed in NCD mice, but also suppressed appetite and raised AKT levels in liver tissues that also required GHSR. Peripheral nesfatin-1 suppressed c-fos expression of GHSR immunoreactive neurons induced by fasting in hypothalamic nuclei, indicating that nesfatin-1 inhibited the activation of central GHSR. We concluded that the effects of nesfatin-1 on food intake and glucose metabolism were GHSR-dependent, and that the glycemic effect was associated with AKT and GLUT4. This study should stimulate further exploration of the nesfatin-1 receptor.