Alterations of Spontaneous Cortical and Subcortical Activity in Type 2 Diabetes Mellitus with and without Depression.

INTRODUCTION: Type 2 diabetes mellitus (T2DM) patients with depression have a higher risk of complications and mortality than T2DM without depression. However, the exact neuropathophysiological mechanism remains unclear. Consequently, the current study aimed to investigate the alteration of cortical and subcortical spontaneous neural activity in T2DM patients with and without depression. METHODS: The demographic data, clinical variables, neuropsychological tests, and functional and anatomical magnetic resonance imaging of depressed T2DM (n = 47) of non-depressed T2DM (n = 59) and healthy controls (n = 41) were collected and evaluated. The correlation analysis, stepwise multiple linear regression, and receiver operating characteristic curve were performed for further analysis. RESULTS: Abnormal neural activities in the bilateral posterior cingulate cortex (PCC) and hippocampus were observed in depressed and non-depressed T2DM and the right putamen of the depressed T2DM. Interestingly, the subcortical degree centrality (DC) of the right hippocampus and putamen were higher in depressed than non-depressed T2DM. Furthermore, the cortical amplitude of low-frequency fluctuation (ALFF) in PCC, subcortical DC in the putamen of depressed T2DM, and hippocampus of non-depressed T2DM was correlated with cognitive scores. In contrast, the cortical fractional ALFF in PCC of non-depressed T2DM was correlated with depression scores. CONCLUSIONS: The abnormalities of spontaneous cortical activity in PCC and subcortical activity in the hippocampus might represent the neurobiological feature of cerebral dysfunction in T2DM. Notably, the altered subcortical activity in the right putamen might mainly associate with negative emotion in T2DM, which could be a promising biomarker for recognizing early cerebral dysfunction in depressed T2DM. This study provided a novel insight into the neuropathophysiological mechanism of brain dysfunction in T2DM with and without depression.

The intestinal absorption mechanism of chicoric acid and its bioavailability improvement with chitosan.

Chicoric acid (CA), an active phenolic acid of Echinacea purpurea (Linn.) Moench, has been demonstrated to exhibit antioxidative, antiviral and immunological activities. A prior study showed that CA is a water-soluble compound with low bioavailability. The current study was performed to study the intestinal absorption mechanism of CA and improve its bioavailability using natural biodegradable chitosan. A Caco-2 monolayer cell model was established to characterise the mechanisms involved in the intestinal absorption of CA. The bioavailability improvement of CA was studied in Sprague-Dawley rats after oral (20 mg/kg) administration of 0.5% chitosan. In vitro, the results showed that the absorption transport of CA was fairly poor, with Papp values of 8.2 × 10-8 to 2.1 × 10-7 cm/s in the absorption direction and 1.5 × 10-7 to 2.6 × 10-7 cm/s in the secretory direction. The permeability was increased by EDTA and chitosan in both directions. Moreover, the transport through the intestinal monolayer was H+ dependent, and P-glycoprotein and OATP2B1 transporters were involved in the intestinal transport of CA. In vivo, the absorption of CA was increased and accelerated with chitosan in rats because the bioavailability was 1.74-fold that of the prototype drug. The above mentioned results indicated that CA was a poor absorption drug and that paracellular and carrier-mediated trancellular transport both participated in its transport route. Chitosan is an excellent absorption enhancer for CA. The transport characteristics uncovered in this study lay the groundwork for further studies directed toward the development and utilisation of its new formulations.

Development of a molecularly imprinted microspheres-based microplate fluorescence method for detection of amantadine and rimantadine in chicken.

In this study, molecularly imprinted microspheres of a type capable of recognising amantadine and rimantadine were first synthesised, and three fluorescent tracers based on dansyl chloride, fluorescein isothiocyanate and 5-carboxytetramethylrhodamine were also synthesised. These reagents were used to develop and optimise a direct competitive fluorescence method on conventional 96-well microplate for detection of the two analytes. Results showed that this method achieved simple operation procedure, rapid assay process (30 min), high sensitivity (limits of detection 0.04-0.05 ng mL-1) and acceptable recycle performance (five times). After optimisation of several parameters, this method was used to detect amantadine and rimantadine in chicken muscle samples. Their recoveries from standards fortified blank samples were in the range of 62.3-93.7%. The analysis results for some real chicken samples were consistent with a confirmatory LC-MS/MS method. Therefore, this method could be used as a rapid, simple and accurate tool for routine screening the residues of amantadine and rimantadine in a large number of chicken muscle samples.

Production of generic monoclonal antibody and development of chemiluminescence immunoassay for determination of 32 sulfonamides in chicken muscle.

A hapten of sulfabenzamide was first synthesized to generate a monoclonal antibody that simultaneously recognized 32 sulfonamides. The computational simulation showed that the 3D conformation, molecular bend angle, molecular volume, electronic charge of core structure of these drugs all showed influences on the antibody binding. The antibody was combined with a heterologous enzyme-labeled hapten to develop a direct competitive chemiluminescence enzyme linked immunosorbent assay for determination of the 32 sulfonamides in chicken muscle sample. The CRs of the optimized method for these drugs were in the range of 7.3%-1778%, and the IC50 values were in the range of 0.038-11.2 ng/g. The limits of detection for detection of these drugs in chicken were in the range of 0.03-26 ng/g. Their recoveries from the standards fortified blank chicken samples were in the range of 60.8%-97.1%. Therefore, this method could be used as a useful tool for routine screening sulfonamides residues in meat.

A microtitre chemiluminescence sensor for detection of pyrethroids based on dual-dummy-template molecularly imprinted polymer and computational simulation.

The residues of pyrethroids in foods of animal origin are dangerous to the consumers, so this study presented a chemiluminescence sensor for determination of pyrethroids in chicken samples. A dual-dummy-template molecularly imprinted polymer capable of recognizing 10 pyrethroids was synthesized. The results of computation simulation showed that the specific 3D conformations of the templates had important influences on the polymer' recognition ability. The polymer was used to prepare a sensor on conventional 96-well microplates, and the sample solution was added into the wells for direct absorption. The absorbed analytes were initiated with the bis(2,4,6-trichlorophenyl)oxalate-H2 O2 -imidazole system, and the chemiluminescence intensity was used for analyte quantification. Results showed that one assay was finished within 12 min, and this sensor could be reused four times. The limits of detection for the 10 analytes were in the range o0.3-6.0 pg/ml, and the recoveries from the standards of fortified blank chicken samples were in the range 70.5-99.7%.

Detection of chloramphenicol in meat with a chemiluminescence resonance energy transfer platform based on molecularly imprinted graphene.

In this study, a novel composite was synthesized by polymerizing the dummy-template molecularly imprinted microspheres on the surface of magnetic graphene. This composite was used as recognition reagent and energy acceptor to develop a platform for determination of chloramphenicol according to the principle of chemiluminescence resonance energy transfer. The light signal was induced with luminolH2O24-(imidazole-1-yl)phenol system, and the chemiluminescence intensity was positively correlated with the analyte concentration. The limit of detection for chloramphenicol in meat sample was 2.0 pg/g, and the recoveries from the standard fortified blank meat sample were in the range of 69.5%-97.3%. Furthermore, one single assay could be finished within 10 min, and the magnetic composite could be reused for at least thirty times. Therefore, this platform could be used as a rapid, simple, sensitive, accurate and recyclable tool for screening the residue of chloramphenicol in meat.

Determination of sulfonamides in meat with dummy-template molecularly imprinted polymer-based chemiluminescence sensor.

In this study, a molecularly imprinted polymer capable of recognizing 15 sulfonamides was first synthesized with sulfabenz as the dummy template. The calculation results from computation simulation showed that the specific 3D conformation of the template had an important influence on the polymer's recognition ability. Then, the polymer was used as recognition reagent to prepare a chemiluminescence sensor on a conventional 96-well microplate for the determination of the residues of 15 sulfonamides in meat (chicken and pork). Due to the 4-(imidazol-1-yl)phenol-enhanced luminol-H2O2 system, the limits of detection for the 15 analytes were in the range of 1.0-12 pg/mL. The recoveries from the standard fortified blank samples were in the range of 72.7-99%. Furthermore, one assay could be finished within 30 min, and the sensor could be reused 4 times. Therefore, this sensor could be used as a very useful tool for routine screening of residues of sulfonamides in meat samples. Graphical abstract Assay procedures of the molecularly imprinted polymer-based chemiluminescence sensor for determination of sulfonamides.

Dummy molecularly imprinted polymer based microplate chemiluminescence sensor for one-step detection of Sudan dyes in egg.

The objective of this study is to report a molecularly imprinted polymer-based chemiluminescence method for determination of Sudan dyes. A dummy-template molecularly imprinted polymer capable of recognizing seven Sudan dyes was first synthesized and its recognition mechanism was studied by using computation simulation method. The polymer was coated in the wells of conventional microplate to prepare a chemiluminescence sensor and the assay process consisted of only one sample-loading step prior to signal acquisition. The optimized sensor was used to determine the seven dyes in egg yolk and the results were confirmed with a high performance liquid chromatography. Results showed that this sensor achieved ultrahigh sensitivity (1.0-5.0 pg/mL), rapid assay process (10 min) and satisfactory recovery (70.5%-92.2%). Furthermore, the sensor could be reused for 5 times. Therefore, this sensor could be used as a useful tool for screening the residues of Sudan dyes in egg.

Detection of chloramphenicol in chicken, pork and fish with a molecularly imprinted polymer-based microtiter chemiluminescence method.

In this study, 4-nitrotoluene (NT) was used as dummy template to synthesize a molecularly imprinted polymer that was highly specific for chloramphenicol. The polymer was coated in the wells of 96-well microplates as recognition reagent to develop a chemiluminescence method. The analyte solution and an enzyme-labelled hapten were added into the wells to perform competition, and the light signal was induced with a highly efficient luminol-H2O2-4-(imidazol-1-yl)phenol system. Then, the optimized method was used to determine chloramphenicol in meat (chicken, pork and fish), and the limit of detection (LOD) was 5.0 pg g-1. Furthermore, the polymer-coated plate could be reused four times, and one test could be finished within 20 min. The recoveries from the standard fortified blank meat samples were in the range of 71.5-94.4%. Therefore, this method could be used as a useful tool for routine screening the residue of chloramphenicol in meat samples.

Application of molecularly imprinted polymer based matrix solid phase dispersion for determination of fluoroquinolones, tetracyclines and sulfonamides in meat.

In this study, a type of novel mixed-template molecularly imprinted polymer was synthesized that was able to recognize 8 fluoroquinolones, 8 sulfonamides and 4 tetracyclines simultaneously with recoveries higher than 92%. Then the polymer was used to develop a matrix solid phase dispersion method for simultaneous extraction of the 20 drugs in pork followed by determination with ultra performance liquid chromatography. During the experiments, the MMIP amount, washing solvent and elution solvent were optimized respectively. The limits of detection of this method for the 20 drugs in pork were in the range of 0.5-3.0ngg-1, and the intra-day and inter-day recoveries from the fortified blank samples were in the range of 74.5%-102.7%. Therefore, this method could be used as a rapid, simple, specific and sensitive method for multi-determination of the residues of the three classes of drugs in meat.

Dummy template molecularly imprinted polymer for solid phase extraction of phenothiazines in meat based on computational simulation.

The 3D structures of two dummy templates and four phenothiazine drugs were studied by using computational simulation method. Then the two dummy templates were used to synthesize two molecularly imprinted polymers respectively. Results showed that the recognition abilities of the two polymers were consistent with the theoretical calculation. Then a solid phase extraction column was developed for extraction of the four phenothiazines in meat (pork, chicken) followed by determination with high performance liquid chromatography. The column showed high adsorption capacities (850-962ng analyte per milligram of polymer) and high recoveries (93-98%) to the four drugs, and could be recycled for sixty times. The limits of detection were in the range of 1.0-10ng/g, and the recoveries from the fortified blank samples were in the range of 70.3-96.1%. This is the first study reporting the use of molecularly imprinted polymer-based method for determination of phenothiazines residues in foods.

Validation of the association between PSMA6 -8 C/G polymorphism and type 2 diabetes mellitus in Chinese Dongxiang and Han populations.

OBJECTIVE: The aim of our study was to validate association between -8 C/G variant of PSMA6 gene and T2DM in Chinese Dongxiang and Han populations. METHOD: We genotyped PSMA6 gene -8 C/G polymorphism in the control groups and T2DM groups in two populations from China using PCR-RFLP technique. Phenotypes and biochemical indicators were measured by biochemical technique. RESULT: The frequencies of CG+GG genotype were observably different from CC genotype in the T2DM groups and control groups (for Dongxiang population: OR = 1.341, 95% CI: 1.101-1.632, P = 0.004; for Han population: OR = 1.313, 95% CI: 1.085-1.569, P = 0.006 after adjusting for gender, age, and BMI, respectively). In the Dongxiang population, the FPG, HOMA-IR, SBP and TG levels of CG+GG genotype were markedly higher than those of the CC genotype in control group (all P < 0.05). However, in the Han population, we only found that the FPI level of the CC genotype was significantly higher than that of the CG+GG genotype in control group (P < 0.05). CONCLUSION: Our investigation suggests that -8 C/G variant of PSMA6 gene may be associated with T2DM and diabetes-related metabolic traits in Chinese Dongxiang and Han populations.

Association of P213S polymorphism of the L-selectin gene with type 2 diabetes and insulin resistance in Chinese population.

AIMS: L-selectin belongs to selectin family of adhesion molecule and participates in the generation and development of type 2 diabetes (T2D). In this study, we evaluated the relationship between the P213S polymorphism of L-selectin gene and T2D and insulin resistance in the Chinese population. METHODS: We genotyped P213S polymorphism in 801 patients with T2D and 834 healthy controls in the Chinese population using polymerase chain reaction-ligase detection reaction (PCR-LDR) technique. Plasma glucose, insulin, lipid, blood urea nitrogen, creatinine and uric acid levels were measured by biochemical technique. RESULTS: The frequency of 213PP genotype and P allele of the L-selectin gene in patients with T2D was significantly higher than that in controls (P=0.007; P=0.019, respectively). The relative risk of allele P suffered from T2D was 1.191 times higher than that of allele S. Moreover, the levels of FPG and HOMA-IR of PP and PS genotype carriers were significantly higher than those of SS genotype carriers in the T2D group (P<0.05). CONCLUSION: These findings indicated that the P213S polymorphism of L-selectin gene may contribute to susceptibility to T2D and insulin resistance in the Chinese population, and P allele appears to be a risk factor for T2D.

[Association of sterol regulatory element binding protein-1c genetic polymorphisms rs2297508 and rs11868035 with type 2 diabetes mellitus in Gansu Han and Dongxiang population].

OBJECTIVE: To assess the association between single nucleotide polymorphisms (SNP) rs2297508 and rs11868035 of sterol regulatory element binding protein-1c (SREBP-1c) gene and type 2 diabetes mellitus (T2DM) in Han and Dongxiang populations in Gansu. METHODS: A total of 342 patients with T2DM and 343 healthy controls of Han ethnics and 218 patients with T2DM and 238 healthy controls of Dongxiang ethnics were enrolled during 2006 and 2009. Genotypes of rs2297508 and rs11868035 were determined by polymerase chain reaction-denaturing high performance liquid chromatography (PCR-DHPLC). Plasma levels of lipid, glucose and insulin in all subjects were assayed by biochemical techniques. Chi-square test was used for data analysis. RESULTS: The genotypic and allelic frequencies of rs2297508 and rs11868035 polymorphisms did not differ significantly in the control groups of Han and Dongxiang ethnics (P> 0.05). But those of T2DM patients were significantly higher than in the controls (P< 0.01). Plasma low density lipoprotein cholesterol (LDL-C) levels for carriers of C allele for rs2297508 were significantly higher than those of non-carriers from the control group of Han ethnics (P<0.05). That of carriers of CC genotypes for rs11868035 was significantly higher than carriers of GG genotype in the control group of Dongxiang ethnics (P<0.05). CONCLUSION: SNP rs2297508 and rs11868035 of SREBP-1c gene may be associated with T2DM, and C allele appears to be a risk factor for T2DM in Gansu Han or Dongxiang ethnics. Han and Dongxiang ethnics did not differ significantly in terms of genotypic and allelic frequencies for the two SNPs. SNP rs2297508 of SREBP-1c gene may be associated with increased plasma levels of LDL-C in both Han or Dongxiang ethnics.

Association of ghrelin Leu72Met polymorphism with type 2 diabetes mellitus in Chinese population.

BACKGROUND: Ghrelin, a novel endogenous ligand for the growth hormone secretagogue receptor, is considered to implicate the development of the type 2 diabetes mellitus (T2DM). The Leu72Met (+408C>A) polymorphism of the preproghrelin, has been linked to obesity, insulin resistance and diabetes. OBJECTIVE: To investigate the distribution of ghrelin gene Leu72Met polymorphism and its association with the type 2 diabetes mellitus in Chinese population. METHODS: We conducted a case-control study on 877 patients with T2DM and 864 controls, which were genotyped by the polymerase chain reaction (PCR) technique, denaturing high performance liquid chromatography (DHPLC) and DNA sequence analysis. Laboratory analyses were carried out in the hospital laboratory. RESULTS: No significant difference in the Leu72Met genotype distributions and allele frequency was observed between type 2 diabetes mellitus and controls (both P>0.05). The polymorphism was not associated with T2DM. However, among the T2DM group, the patients carrying Leu72Leu genotype had significantly increased levels of FPG and serum creatinine compared with variant genotypes (Leu72Met and Met72Met) (P<0.05). In the control group, the subjects with variant genotypes had significantly increased levels of FINS, HOMA-IR compared with Leu72Leu genotype (P<0.05). CONCLUSION: The Leu72Met polymorphism of the preproghrelin gene was not associated with T2DM in Chinese population. However, it may have some roles in the etiology of insulin resistance.

Multidetermination of four nitrofurans in animal feeds by a sensitive and simple enzyme-linked immunosorbent assay.

In this study, the polyclonal antibody against furazolidone was produced with furazolidone coupling to protein carriers by a diazotization method and glutaraldehyde reaction, respectively. The antibody obtained showed good specificity toward furazolidone and various cross-reactivity toward nitrofurantoin, nitrofurazone, and furaltadone. Then, an indirect competitive enzyme-linked immunosorbent assay (ELISA) based on the antibody was first developed for multidetermination of four nitrofurans in animal feeds. The limit of detection (LOD) of the method was 0.2-2.1 ng/g depending on the component. After simple extraction, the fortified swine and broiler chicken feed samples were detected with recovery ranges of 75.9-86.4%. Results obtained from ELISA were confirmed by high-performance liquid chromatography (HPLC) with ultraviolet detection. Analysis of the unknown feed samples indicates that ELISA can be a practical tool for screening of nitrofurans in animal feeds before confirmation by HPLC.

Evaluation of the association between retinal binding protein 4 polymorphisms and type 2 diabetes in Chinese by DHPLC.

Serum retinal binding protein 4 (RBP4) was recently described as a new liver- and adipocyte-derived signal that may contribute to Type 2 diabetes mellitus (T2DM) and insulin resistance. The aim of this study was to test whether the RBP4 gene could be used as a genetic marker to predict the development of T2DM amongst the Chinese population of Han. For this study, a normal control group of 115 healthy subjects and an experimental group of 107 patients with T2DM were examined. A combined method of denaturing high-performance liquid chromatography (DHPLC) and sequencing was applied to the detection of the RBP4 gene variants. Two SNPs, rs17484721 and rs36035572, were analyzed. Phenotypes and biochemical indicators related to the metabolism of glucose and lipid were measured. We found that there are significant differences between the control group and the patients group in terms of their respective distributions of genotype and allele frequency. The TG levels of the TT and II genotype was significantly higher than that of the TC + CC and ID + DD, respectively, in both patient group and control group. These findings suggest that the variations in the RBP4 gene may be associated with T2DM and serum triglyeride levels in the Han Chinese.

Association of sterol regulatory element-binding protein-1c gene polymorphism with type 2 diabetes mellitus, insulin resistance and blood lipid levels in Chinese population.

AIMS: The sterol regulatory element-binding protein (SREBP)-1c gene has been identified as a susceptibility gene in metabolic diseases such as type 2 diabetes mellitus (T2DM), obesity, dyslipidemia and insulin resistance. Previous studies suggest that SNP17 (rs2297508, exon18c and G952G) of SREBP-1c gene and a common SREBP-1c SNP6 (rs11868035) are associated with an increased risk of T2DM. The present study aimed to confirm the previously reported association in a Chinese population and to examine the two SREBP-1c SNPs for their associations with insulin resistance and blood lipid. METHODS: We genotyped two SREBP-1c SNPs in a case-control study (n=327) from Chinese, including 156 patients with T2DM and 171 healthy controls, using polymerase chain reaction-denaturing high-performance liquid chromatography (PCR-DHPLC) and tested for association with type 2 diabetes, insulin resistance and blood lipid, respectively. Genotype and allele distributions and haplotype construction were analysed. RESULTS: The genotype and allele distributions of rs2297508 and rs11868035 polymorphisms were significantly different in type 2 diabetic patients compared to controls (P=0.002 and P=0.013; 0.00 and 0.001, respectively). Haplotype analyses showed significant association with diabetes risk and confirmed the results of the single SNP analyses. The plasma levels of LDL-c of the minor allele-C carriers of the two SNPs were both significantly higher than the noncarriers in the control group (P<0.05). Furthermore, insulin resistance index (HOMA-IRI) of the rare homozygotes C/C of rs11868035 was significantly lower than that of T/T in the T2DM group (P<0.05). CONCLUSIONS: These findings indicate that the SREBP-1c SNPs rs2297508 and rs11868035 are associated with a significantly increased risk of T2DM and dyslipidemia in the Chinese population. Moreover, the SNP (rs11868035) is closely related to insulin resistance (IR) in diabetic patients.