MdbHLH4 negatively regulates apple cold tolerance by inhibiting MdCBF1/3 expression and promoting MdCAX3L-2 expression.

Low temperature affects the yield and quality of crops. Inducer of CBF expression 1 (ICE1) plays a positive role in plant cold tolerance by promoting the expression of CRT binding factor (CBF) and cold-responsive (COR) genes. Several ICE1-interacting transcription factors (TFs) that regulate plant cold tolerance have been identified. However, how these TFs affect the function of ICE1 and CBF expression under cold conditions remains unclear. Here, we identified the MYC-type TF MdbHLH4, a negative regulator of cold tolerance in Arabidopsis (Arabidopsis thaliana) and apple (Malus domestica) plants. Under cold conditions, MdbHLH4 inhibits the expression of MdCBF1 and MdCBF3 by directly binding to their promoters. It also interacts with MdICE1L, a homolog of AtICE1 in apple, and inhibits the binding of MdICE1L to the promoters of MdCBF1/3 and thus their expression. We showed that MdCAX3L-2, a Ca2+/H+ exchanger (CAX) family gene that negatively regulates plant cold tolerance, is also a direct target of MdbHLH4. MdbHLH4 reduced apple cold tolerance by promoting MdCAX3L-2 expression. Moreover, overexpression of either MdCAX3L-2 or MdbHLH4 promoted the cold-induced ubiquitination and degradation of MdICE1L. Overall, our results reveal that MdbHLH4 negatively regulates plant cold tolerance by inhibiting MdCBF1/3 expression and MdICE1L promoter-binding activity, as well as by promoting MdCAX3L-2 expression and cold-induced MdICE1L degradation. These findings provide insights into the mechanisms by which ICE1-interacting TFs regulate CBF expression and ICE1 function and thus plant cold tolerance.

Iron Metabolism Markers and Lower Extremity Arterial Disease in People with Type 2 Diabetes.

Objective: To determine the levels of serum iron, ferritin, total iron-binding capacity, and hepcidin in patients with type 2 diabetes mellitus (T2DM), and to elucidate the relationship of these biomarkers with lower extremity arterial disease (LEAD). Methods: Three hundred fifteen patients with T2DM were selected for the study and divided into non-LEAD (n = 119) and LEAD groups (n=196) based on the ankle-brachial index (ABI) results. Demographic data and clinical test results were collected from all patients. Serum iron, ferritin, total iron-binding capacity, and hepcidin levels were measured, and the transferrin saturation was calculated. Results: Hepcidin levels were substantially higher in the LEAD group (19.17 ± 8.66 ng/mL) than the non-LEAD group (15.44±7.55 ng/mL, P < 0.001), and there was a negative correlation between the ABI and serum lecithin level (r = -0.349, P < 0.001). There were no other correlations with the other iron metabolism indicators. The results of dichotomous logistic regression with LEAD as the dependent variable revealed that smoking history (OR = 4.442, P = 0.008), hypertension history (OR = 3.721, P = 0.006), cardiovascular disease history (OR = 11.126, P < 0.001), diabetes duration (OR = 1.305, P < 0.001), age (OR = 1.056, P = 0.021), hs-CRP level (OR = 1.376, P = 0.002), HbA1c concentration (OR = 1.394, P = 0.001), and hepcidin level (OR = 1.097, P = 0.003) were independent risk factors for LEAD in T2DM patients. Conclusion: Serum hepcidin levels were elevated in the LEAD group compared with the non-LEAD group, and elevated hepcidin levels were associated with the development of LEAD in T2DM patients, suggesting that hepcidin may be involved in the occurrence and development of LEAD in T2DM patients.

The Annotation of Zebrafish Enhancer Trap Lines Generated with PB Transposon.

An enhancer trap (ET) mediated by a transposon is an effective method for functional gene research. Here, an ET system based on a PB transposon that carries a mini Krt4 promoter (the keratin4 minimal promoter from zebrafish) and the green fluorescent protein gene (GFP) has been used to produce zebrafish ET lines. One enhancer trap line with eye-specific expression GFP named EYE was used to identify the trapped enhancers and genes. Firstly, GFP showed a temporal and spatial expression pattern with whole-embryo expression at 6, 12, and 24 hpf stages and eye-specific expression from 2 to 7 dpf. Then, the genome insertion sites were detected by splinkerette PCR (spPCR). The Krt4-GFP was inserted into the fourth intron of the gene itgav (integrin, alpha V) in chromosome 9 of the zebrafish genome, with the GFP direction the same as that of the itgav gene. By the alignment of homologous gene sequences in different species, three predicted endogenous enhancers were obtained. The trapped endogenous gene itgav, whose overexpression is related to hepatocellular carcinoma, showed a similar expression pattern as GFP detected by in situ hybridization, which suggested that GFP and itgav were possibly regulated by the same enhancers. In short, the zebrafish enhancer trap lines generated by the PB transposon-mediated enhancer trap technology in this study were valuable resources as visual markers to study the regulators and genes. This work provides an efficient method to identify and isolate tissue-specific enhancer sequences.

Genome-Wide Analysis of the Apple CBL Family Reveals That Mdcbl10.1 Functions Positively in Modulating Apple Salt Tolerance.

Abiotic stresses are increasingly harmful to crop yield and quality. Calcium and its signaling pathway play an important role in modulating plant stress tolerance. As specific Ca2+ sensors, calcineurin B-like (CBL) proteins play vital roles in plant stress response and calcium signaling. The CBL family has been identified in many plant species; however, the characterization of the CBL family and the functional study of apple MdCBL proteins in salt response have yet to be conducted in apple. In this study, 11 MdCBL genes were identified from the apple genome. The coding sequences of these MdCBL genes were cloned, and the gene structure and conserved motifs were analyzed in detail. The phylogenetic analysis indicated that these MdCBL proteins could be divided into four groups. The functional identification in Na+-sensitive yeast mutant showed that the overexpression of seven MdCBL genes could confer enhanced salt stress resistance in transgenic yeast. The function of MdCBL10.1 in regulating salt tolerance was also verified in cisgenic apple calli and apple plants. These results provided valuable insights for future research examining the function and mechanism of CBL proteins in regulating apple salt tolerance.

Cation/Ca2+ Exchanger 1 (MdCCX1), a Plasma Membrane-Localized Na+ Transporter, Enhances Plant Salt Tolerance by Inhibiting Excessive Accumulation of Na+ and Reactive Oxygen Species.

High salinity causes severe damage to plant growth and significantly reduces crop yields. The CCX family proteins can facilitate the transport of multiple ions to prevent toxicity. CCX proteins play an important role in regulating plant salt tolerance, but no detailed studies on CCX proteins in apples have been reported. Here, the CCX family gene MdCCX1 was cloned from apple (Malus domestica). It is constitutively expressed in various apple tissues and is significantly induced by salt stress. As a plasma membrane-localized protein, MdCCX1-overexpression could complement the Na+-sensitive phenotype of yeast mutants and reduce the Na+ content in yeast cells under NaCl treatment, suggesting that MdCCX1 could be a plasma membrane-localized Na+ transporter. To identify the function of MdCCX1 in salt response, we transformed this gene into Arabidopsis, apple calli, and apple plants. Overexpression of MdCCX1 significantly improved the salt tolerance of these transgenic materials. The significantly reduced Na+ content under NaCl treatment indicated that MdCCX1 overexpression could enhance plant salt tolerance by inhibiting the excessive accumulation of Na+. Besides, MdCCX1 overexpression could also enhance plant salt tolerance by promoting ROS scavenging. These findings provide new insight and rich resources for future studies of CCX proteins in plant species.

The Influence of Interpersonal Sensitivity on Smartphone Addiction: A Moderated Mediation Model.

Smartphone addiction is a behavioral dependence characterized by excessive or compulsive Internet use and a preoccupation with and loss of control over this use that interferes with an individual's daily functioning and results in negative mental processes and subsequent social consequences. Smartphone addiction can negatively impact physical and mental health as well as academic performance, sleep quality, and even interpersonal interaction and relationships. Based on the compensatory Internet use theory, this study explores the relationship between interpersonal sensitivity and smartphone addiction in college students and constructed a moderated mediation model. A sample of 881 college students was tested using the Interpersonal Sensitivity Scale, Smart Phone Addiction Scale, Fear of Missing Out Scale, and Relational Self-Construal Scale. We used AMOS 26.0 to conduct a confirmatory factor analysis and employed SPSS 24.0 to test our hypotheses. The results indicated that (1) interpersonal sensitivity was positively related to the fear of missing out and smartphone addiction; (2) the fear of missing out mediated the relationship between interpersonal sensitivity and mobile phone addiction; (3) relational self-construal moderated interpersonal sensitivity and the fear of missing out; and (4) relational self-construal moderated the mediating effect of the fear of missing out on the relationship between interpersonal sensitivity and smartphone addiction. We concluded that the fear of missing out and relational self-construal play a moderated mediation effect on the relationship between smartphone addiction and interpersonal sensitivity. Our findings provided some theoretical implications. Specifically, in addition to proposing a new approach for the study of smartphone addiction, we also introduced a theoretical basis for psychotherapy and intervention of smartphone addiction. In addition, this study also provides some insightful ideas for educational practitioners.

Preparation of clarithromycin floating core-shell systems (CSS) using multi-nozzle semi-solid extrusion-based 3D printing.

Matrix erosion is unavoidable during the release of poorly soluble drugs from gastric floating delivery system (GFDDS), which shortens the floating time and diminishes drug release. We fabricated a core-shell system (CSS) consisting of a low-density drug-loaded shell and a floating core using multi-nozzle semi-solid extrusion (SSE) 3D printing technology. The clarithromycin (CAM) loading capacity of the shell was 81.7%. The floating core paste provided structural support during printing and formed a hollow structure in CAM CSS, which increased the buoyancy in the early stage of drug release. In addition, the floating core had numerous micro-airbags that swelled when the solution penetrated the core, and generated CO2. The micro-airbag structure and CO2 generation further increased the buoyancy of CSS. The CAM CSS achieved 74.5% (w/w) drug loading, 8 h sustained release, and immediate and prolonged floating (>10 h). This structure of CSS and floating core provide a novel perspective for constructing a stable gastric floating drug delivery system.

Preparation of High-Drug-Loaded Clarithromycin Gastric-Floating Sustained-Release Tablets Using 3D Printing.

The high-drug-loaded sustained-release gastric-floating clarithromycin (CAM) tablets were proposed and manufactured via semisolid extrusion (SSE)-based 3D printing. The physical and mechanical properties, such as dimensions, weight variation, friability, and hardness, were accessed according to the quality standards of Chinese Pharmacopoeia (Ch.P). The interactions among the drug-excipients were evaluated via differential scanning calorimetry (DSC), Fourier-transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) techniques. Next, the rheological properties of the paste and the effect of the excipients and solvents were evaluated. Finally, a very high drug-loading of up to 81.7% (w/w) with the sustain release time of 8 h (125 mg) and 12 h (250 mg) was achieved. The results revealed the potential of SSE for achieving a high drug loading and identified the suitable properties of the paste for SSE-based 3D printing.

Serum growth differentiation factor 15 is closely associated with metabolic abnormalities in Chinese pregnant women.

AIMS: To explore the relationship between serum growth differentiation factor 15 (GDF15) and metabolic abnormalities in Chinese pregnant women. MATERIALS AND METHODS: We recruited 200 patients with gestational diabetes mellitus (GDM) and 211 matched normal control within 24-28 weeks of pregnancy. Enzyme-linked immunosorbent assay (ELISA) was used to determine the serum GDF15 levels of all participants. Then we grouped participants according to the number of metabolic abnormalities (including blood glucose, blood lipids and blood pressure), divided them into a normal metabolic group, one metabolic abnormality group, two or more metabolic abnormalities group. Finally, multinomial logistic regression analysis was used to estimate the odds radio (OR) and 95% CIs expressing the association between GDF15 and metabolic abnormalities in pregnant women. RESULTS: Through bivariate correlation analysis, we found that serum GDF15 is linearly correlated with glucose metabolism indices, such as 1h-PG, 2h-PG, HbA1c (all P < 0.05). In addition, serum GDF15 and triglycerides were linearly correlated (P < 0.05). Grouping by the number of metabolic abnormalities, we found that as GDF15 levels increased, the risk of metabolic abnormalities also increased (OR > 1), and the risk of multiple metabolic abnormalities was higher. As the number of metabolic abnormalities increased, serum GDF15 levels also were elevated (P < 0.001). CONCLUSIONS: The results suggest that serum GDF15 levels are closely associated with metabolic abnormalities in pregnant women and may be used as a predictor of metabolic abnormalities during pregnancy.

miR­135b­5p enhances the sensitivity of HER­2 positive breast cancer to trastuzumab via binding to cyclin D2.

Trastuzumab has led to a marked improvement in the outcomes of patients with human epidermal growth factor receptor 2 (HER­2)­positive breast cancer. However, the effects of trastuzumab on HER­2­positive breast cancer are limited by the emergence of its cardiotoxicside effects. MicroRNA (miR)­135b­5p has been shown to inhibit tumor metastasis in breast cancer. The present study aimed to explore the effects of miR­135b­5p overexpression on the efficacy of trastuzumab in HER­2­positive breast cancer. Reverse transcription­quantitative PCR was performed to detect the levels of miR­135b­5p. Cell viability was evaluated with a Cell Counting Kit­8 assay. Annexin V/propidium iodide staining was employed to detect the number of apoptotic cells. Flow cytometry assay was performed to investigate the cell cycle. Western blotting was used to detect the expression levels of Bax, cleaved caspase­3, Bcl­2, cyclin D2, p27Kip1 and cyclin E1. Cell migration and invasion were detected by Transwell assay. Luciferase assays were conducted to identify the target gene of miR­135b­5p. In addition, an in vivo tumor xenograft model was established. miR­135b­5p agomir significantly enhanced the anti­proliferative effect of trastuzumab on HER­2­positive breast cancer cells via the induction of apoptosis, whereas the anti­metastatic effect of trastuzumab was enhanced by miR­135b­5p agomir treatment. Subsequently, luciferase assays indicated that cyclin D2 was the direct target of miR­135b­5p, whereas overexpression of the latter arrested cell cycleduring the G0/G1 phase. Moreover, miR­135b­5p agomir notably increased the antitumor effect of trastuzumab in vivo. The data demonstrated that miR­135b­5p sensitized HER­2­positive breast cancer cells to trastuzumab in vitro and in vivo by directly binding to cyclin D2. These results suggested that the combination of miR­135b­5p with trastuzumab may be a therapeutic strategy for patients with HER­2­positive breast cancer.

Serum 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid is associated with lipid profiles and might protect against non-alcoholic fatty liver disease in Chinese individuals.

AIMS/INTRODUCTION: High plasma 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF) levels are significantly associated with type 2 diabetes mellitus, which is usually accompanied by metabolic syndrome and non-alcoholic fatty liver disease (NAFLD) with increased triglyceride levels. Thus, we hypothesized that elevated CMPF levels might be related to lipid metabolism and NAFLD risk. MATERIALS AND METHODS: Serum CMPF levels were determined using an enzyme-linked immunosorbent assay in a total of 466 individuals, including 116 controls with no NAFLD or type 2 diabetes mellitus, 53 individuals with NAFLD but no type 2 diabetes mellitus, 151 individuals with type 2 diabetes mellitus but no NAFLD, and 146 individuals with both NAFLD and type 2 diabetes mellitus. The associations with age, blood pressure, lipid profiles, body mass index and liver injury marker levels were examined, and a meta-analysis of non-diabetic and diabetic groups was carried out to detect the combined effects. RESULTS: The CMPF concentration in NAFLD patients was significantly lower than individuals without NAFLD in both the non-diabetic group (P < 0.05) and diabetic group (P < 0.01), and correlated negatively with several parameters of liver function and the adiposity index. Meta-analysis showed that serum CMPF levels was associated with decreased risk of NAFLD after combining the results (odds ratio 0.677, 95% confidence interval 0.552-0.831, P < 0.001). Additionally, the CMPF concentration was independently negatively associated with triglycerides and high-density lipoprotein cholesterol in the meta-analysis. Multiple stepwise regression analysis showed that body mass index, high-density lipoprotein cholesterol, triglyceride level, age, sex and fasting plasma glucose were independently associated with CMPF (all P < 0.05). CONCLUSIONS: The results suggest that serum CMPF levels are negatively related to lipid metabolism and could be used to predict NAFLD development.

Combined effects of octreotide and cisplatin on the proliferation of side population cells from anaplastic thyroid cancer cell lines.

Anaplastic thyroid cancer (ATC) represents the most aggressive subtype of thyroid cancer and has a poor prognosis. In addition to surgery, chemotherapy is an important treatment for ATC; however, the therapeutic effects of current chemotherapies for ATC are not particularly promising. There is a high proportion of side population (SP) cells in ATC, which may be a reason for its drug resistance. In the present study, the antitumor activities of combined octreotide (OCT) and cisplatin (DDP) on the proliferation and apoptosis of ATC SP cells were evaluated. First, SP cells from 8305C and BHT101 cell lines were detected and sorted. Following in vitro culture for 1 week, cluster of differentiation (CD)44, CD133, ATP-binding cassette (ABC) subfamily B member 1 (ABCB1), ABC subfamily G member 2 (ABCG2) and somatostatin receptor expression was detected to characterize the SP cells. An MTT assay was performed to investigate the combined effects on 8305C-SP cell proliferation in vitro, and a mouse model was used to investigate the combined effects on 8305C-SP cell proliferation in vivo. Annexin V/propidium iodide staining was used to investigate the combined effects on 8305C-SP cell apoptosis. Chemotherapeutic drug resistance-associated protein expression and apoptosis-associated protein expression were also detected following combined treatment. As a result, SP cells were identified in 8305C and BHT101 cells, and the proportion of 8305C-SP cells was increased compared with that of BTH101-SP cells. SP cells have enhanced proliferation, tumorigenicity and drug resistance compared with main population cells. The combined treatment of OCT with DDP suppressed the proliferation of 8305C-SP cells in vitro and in vivo, and induced 8305C-SP cell apoptosis. Combined treatment decreased the ABCB1 and ABCG2 expression by SP cells and activated mitochondrial apoptotic signaling, resulting in cell apoptosis. In conclusion, these data support the hypothesis that combined treatment with OCT and DDP induces ATC cell apoptosis and suppresses cell proliferation. These data provide a theoretical basis for further combined chemotherapy clinical applications.

Selective extraction of theophylline from plasma by copper-doped magnetic microspheres prior to its quantification by HPLC.

The authors describe the preparation of copper-doped magnetic microspheres (Cu-Fe3O4) by a solvothermal method. Due to their good magnetic property and high affinity for compounds containing an imidazole moiety (containing N-H), they are excellent adsorbents for such compounds as tested by eighteen compounds. Specifically, a method has been developed for magnetic solid-phase extraction (MSPE) of theophylline (TP) from plasma. The method enables selective enrichment of TP over many potential interferents that can occur in plasma. Following elution with alkaline methanol, TP was quantified by HPLC-UV at a detection wavelength of 272 nm. Under the optimized conditions, a linear response is found for the 0.02 to 20 µg·mL-1 concentration range, and the limit of detection is as low as 3 ng·mL-1. Recoveries from spiked samples range from 91.2 to 100.4%, and the repeatabilities are between 2.9 and 12% (for n = 6). The method was successfully applied to the determination of TP in rabbit and rat plasma. Graphical abstract Copper-doped magnetic microspheres are described that show good magnetic property and high affinity for compounds containing an imidazole moiety (containing an N-H group). They were successfully applied to the selective extraction of theophylline in plasma.

Interaction between Saikosaponin D, Paeoniflorin, and Human Serum Albumin.

Saikosaponin D (SSD) and paeoniflorin (PF) are the major active constituents of Bupleuri Radix and Paeonia lactiflora Pall, respectively, and have been widely used in China to treat liver and other diseases for many centuries. We explored the binding of SSD/PF to human serum albumin (HSA) by using fluorospectrophotometry, circular dichroism (CD) and molecular docking. Both SSD and PF produced a conformational change in HSA. Fluorescence quenching was accompanied by a blue shift in the fluorescence spectra. Co-binding of PF and SSD also induced quenching and a conformational change in HSA. The Stern-Volmer equation showed that quenching was dominated by static quenching. The binding constant for ternary interaction was below that for binary interaction. Site-competitive experiments demonstrated that SSD/PF bound to site I (subdomain IIA) and site II (subdomain IIIA) in HSA. Analysis of thermodynamic parameters indicated that hydrogen bonding and van der Waals forces were mostly responsible for the binary association. Also, there was energy transfer upon binary interaction. Molecular docking supported the experimental findings in conformation, binding sites and binding forces.

Circulating 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF) levels are associated with hyperglycemia and ß cell dysfunction in a Chinese population.

Several recent clinical studies have suggested that the levels of circulating 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF) are significantly higher in patients with gestational diabetes mellitus (GDM), impaired glucose tolerance (IGT), and type 2 diabetes mellitus (T2DM). This study recruited a total of 516 participants. The following patient populations were enrolled: 99 newly diagnosed cases with T2DM, 219 cases with prediabetes [82 with isolated impaired glucose tolerance (I - IGT), 66 with isolated impaired fasting glucose (I - IFG) and 71 with impaired glucose tolerance and impaired fasting glucose (IGT + IFG)], and 198 cases with normal glucose tolerance [NGT, including 99 first-degree relatives of type 2 diabetes patients (FDRs) and 99 non-FDRs]. We investigated the circulating CMPF levels in subjects with different glucose metabolism statuses and examined the potential link between CMPF and ß cell function. Our results indicate that the serum CMPF levels were elevated in the prediabetes, T2DM, and FDRs groups compared to the NGT group. Additionally, the serum CMPF concentrations were independently and negatively associated with the triglyceride levels and Stumvoll first-phase insulin secretion index. Cumulatively, our findings suggest that the circulating CMPF levels can predict glycolipid metabolism disorders. Furthermore, elevated serum CMPF concentrations may determine hyperglycemia and ß cell dysfunction.

Organic Solar Cells with Controlled Nanostructures Based on Microphase Separation of Fullerene-Attached Thiophene-Selenophene Heteroblock Copolymers.

Heteroblock copolymers consisting of poly(3-hexylthiophene) and fullerene-attached poly(3-alkylselenophene) (T-b-Se-PCBP) were synthesized for organic photovoltaic applications by quasi-living catalyst transfer polycondensation and subsequent conversion reactions. Characterization of the polymers confirmed the formation of well-defined diblock structures with high loading of the fullerene at the side chain (∼40 wt %). Heteroblock copolymer cast as a thin film showed a clear microphase-separated nanostructure approximately 30 nm in repeating unit after thermal annealing, which is identical to the microphase-separated nanostructure of diblock copolymer consisting of poly(3-hexylthiophene) and fullerene-attached poly(3-alkylthiophene) (T-b-T-PCBP). These heteroblock copolymers provide an ideal platform for investigating the effects of nanostructures and interfacial energetics on the performance of organic photovoltaic devices.

Serum Magnesium Concentration Is Inversely Associated with Albuminuria and Retinopathy among Patients with Diabetes.

Aim. To investigate the association between serum magnesium levels and microvascular complications among patients with diabetes. Methods. Patients with diabetes were recruited between April 2012 and January 2015. All patients received an assay of serum magnesium concentration, were screened for 24 h albumin excretion rate, and underwent nonmydriatic fundus photography. Albuminuria and retinopathy were defined accordingly. A total of 3,100 patients with normal serum magnesium levels were included in this study. Results. Patients with albuminuria and/or retinopathy had lower levels of serum magnesium than patients without these complications (P < 0.001). The prevalence of isolated albuminuria, isolated retinopathy, and combined albuminuria and retinopathy decreased as the concentration of serum magnesium increased. Multiple logistic regression analysis indicated that the odds ratio for isolated albuminuria, isolated retinopathy, and concomitant albuminuria and retinopathy decreased by approximately 20% for every 0.1 mmol/L increase in serum magnesium concentration. Conclusion. Serum magnesium levels were negatively associated with the risk of diabetic microvascular complications among patients with serum magnesium levels within the normal range.

Zirconium-doped magnetic microspheres for the selective enrichment of cis-diol-containing ribonucleosides.

Zirconium-doped magnetic microspheres (Zr-Fe3O4) for the selective enrichment of cis-diol-containing biomolecules were easily synthesized via a one-step hydrothermal method. Characterization of the microspheres revealed that zirconium was successfully doped into the lattice of Fe3O4 at a doping level of 4.0 at%. Zr-Fe3O4 possessed good magnetic properties and high specificity towards cis-diol molecules, as shown using 28 compounds. For ribonucleosides, the adsorbent not only has favorable anti-interferential abilities but also has a high adsorption capacity up to 159.4µmol/g. As an example of a real application, four ribonucleosides in urine were efficiently enriched and detected via magnetic solid-phase extraction coupled with high-performance liquid chromatography. Under the optimized extraction conditions, the detection limits were determined to be between 0.005 and 0.017µg/mL, and the linearities ranged from 0.02 to 5.00µg/mL (R≥0.996) for these analytes. The accuracy of the analytical method was examined by studying the relative recoveries of the analytes in real urine samples, with recoveries varying from 77.8% to 119.6% (RSDs<10.6%, n=6). The results indicate that Zr-Fe3O4 is a suitable adsorbent for the analysis of cis-diol-containing biomolecules in practical applications.

Circulating Adipocyte Fatty Acid Binding Protein (FABP4) Levels Are Associated with Irisin in the Middle-Aged General Chinese Population.

BACKGROUND: Adipocyte fatty acid binding protein (FABP4) has been recently characterized as an adipokine that is closely associated with obesity and metabolic syndrome. Irisin, a novel myokine, activates thermogenesis by increasing the transformation of white adipocytes to brown, and it has improved glucose homeostasis in animal models. In this study, we aimed to explore the relationship between serum FABP4 and irisin in middle-aged Chinese subjects. METHODS: A total of 111 normal residents (56 men and 55 women) of Fengxian District who were 40 to 60 years of age were recruited. Circulating FABP4 and irisin were determined by enzyme-linked immunosorbent assay. Anthropometric parameters, oral glucose tolerance test results, hemoglobin A1C (HbA1C), blood lipids, homeostasis model assessment of insulin resistance, homeostasis model assessment-ß and body fat composition were also determined. RESULTS: All participants were categorized by FABP4 tertiles. There were significant differences in blood pressure, body fat percentage, 2-h plasma glucose, and skeletal muscle mass among the three groups (P<0.05). Furthermore, FABP4 levels in the women were significantly higher than in the men (P<0.05). However, there was no sexual dimorphism in serum irisin (P>0.05). To exclude the effect of sex difference, partial correlations analysis showed that FABP4 was positively correlated with diastolic blood pressure (P<0.05) and body fat percentage (P<0.05) negatively correlated with skeletal muscle mass (P<0.05) and irisin (P<0.05), while irisin was positively correlated with HbA1c (P<0.05) and negatively correlated with creatinine (P<0.05). Multivariate regression analysis demonstrated that serum FABP4 was independently associated with skeletal muscle mass (P<0.001), diastolic blood pressure (P<0.05) and irisin (P<0.05) after adjustment for age, body mass index, body fat percentage, total cholesterol and HbA1C. CONCLUSIONS: Elevated FABP4 levels increase the risks of obesity-related metabolic disorders and hypertension. Serum irisin might exert antagonistic effects on FABP4 in the middle-aged Chinese population.

Optimized Extraction of Polysaccharides from Grateloupia livida (Harv.) Yamada and Biological Activities.

Polysaccharides from Grateloupia livida (Harv.) Yamada (GL) were extracted by a heating circumfluence method. Single-factor experiments were performed for the three parameters: extraction time (X1), extraction temperature (X2) and the ratio of water to raw material (X3) and their test range. From preliminary experimental results, one type of the response surface methodology, the Box-Behnken design was applied for the optimizing polysaccharide extraction conditions. The experimental data obtained were fitted to a second-order polynomial equation. The optimal conditions were extraction time 5 h, extraction temperature 100 °C and ratio of water to raw material 70 mL/g. Under these conditions, the experimental yield was 39.22% ± 0.09%, which well matched the predicted value (39.25%), with 0.9774 coefficient of determination (R²). GL polysaccharides had scavenging activities for DPPH and hydroxyl radicals in vitro. The scavenging rates for both radicals peaked at 20 mg/mL GL concentration. However, the positive standard, VC (ascorbic acid), possessed stronger antioxidant activities than GL polysaccharides. Furthermore, the anticancer activity of GL polysaccharides on HepG2 cell proliferation increased dose- and time-dependently, but the positive standard, 5-fluorouracil (5-fu) showed more significant anticancer activity in this study. Overall, GL polysaccharides may have potential applications in the medical and food industries.