BACKGROUND/AIMS: Icodextrin (glucose polymer) is metabolized by a-amylase to oligosaccharides such as maltose and maltotriose. The presence of these metabolites could have an effect on the enzymatic glucose measurement especially the glucose dehydrogenase pyrroloquinolinequinone (GDH-PQQ) based method. Patients treated with icodextrin are at risk for inaccurate blood glucose measurements. In this study we measured the blood glucose with different methods and analyzed the results to determine the test accuracy. METHODS: The blood glucose was measured, in seven outpatients and in seven inpatients using icodextrin, by the glucose hexokinase laboratory technique method as well as the GDH-PQQ method (Accu Chek Active)at the same time. To estimate an icodextrin residual effect, after discontinuing icodextin, the blood glucose was measured by the two methods after 48 hours in 4 inpatients. RESULTS: In seven outpatients the blood glucose was overestimated by the Accu Chek Active method (mean difference 68 mg/dL, p value 0.012). In seven inpatients the mean difference in the glucose was 56 mg/dL at 6am, 52 mg/dL at 11am, 52 mg/dL at 4pm, and 50 mg/dL at 9pm by the two different methods. In the four inpatients after changing their dialysate, the mean difference in the glucose was 58 mg/dL after 10 hours, 45 mg/dL after 24 hours, 24 mg/dL after 34 hours, and 26 mg/dL after 48 hours. CONCLUSION: Blood glucose was overestimated by the GDH-PQQ method and the inaccuracies were observed for more than 48 hours.
Blood Glucose ; Glucans ; Glucose ; Glucose 1-Dehydrogenase ; Hexokinase ; Humans ; Hypoglycemia ; Inpatients ; Maltose ; Oligosaccharides ; Outpatients ; Peritoneal Dialysis, Continuous Ambulatory ; Trisaccharides

OBJECTIVE: To investigate whether autophagy mediates the effects of aldehyde dehydrogenase 2 (ALDH2) on the proliferation of neonatal rat cardiac fibroblasts cultured in high glucose. METHODS: Cardiac fibroblasts were isolated from neonatal (within 3 days) SD rats and subcultured. The fibroblasts of the third passage, after identification with immunofluorescence staining for vimentin, were treated with 5.5 mmol/L glucose (control group), 30 mmol/L glucose (high glucose group), or 30 mmol/L glucose in the presence of Alda-1 (an ALDH2 agonist), daidzin (an ALDH2 2 inhibitor), or both. Western blotting was employed to detect ALDH2, microtubule-associated protein 1 light chain 3B subunit (LC3B) and Beclin-1 in the cells, and a hydroxyproline detection kit was used for determining hydroxyproline content in cell culture medium; CCK- 8 kit was used for assessing the proliferation ability of the cardiac fibroblasts after the treatments. RESULTS: Compared with the control cells, the cells exposed to high glucose exhibited obviously decreased expressions of ALDH2, Beclin-1 and LC3B and increased cell number and hydroxyproline content in the culture medium. Treatment of the high glucose-exposed cells with Alda-1 significantly increased Beclin-1, LC3B, and ALDH2 protein expressions and lowered the cell number and intracellular hydroxyproline content, whereas the application of daidzin resulted in reverse changes in the expressions of ALDH2, Beclin-1 and LC3B, viable cell number and intracellular hydroxyproline content in high glucose-exposed cells. CONCLUSIONS Mitochondrial ALDH2 inhibits the proliferation of neonatal rat cardiac fibroblasts induced by high glucose, and the effect is possibly mediated by the up-regulation of autophagy-related proteins Beclin-1 and LC3B.
Aldehyde Dehydrogenase ; Aldehyde Dehydrogenase, Mitochondrial ; metabolism ; Animals ; Animals, Newborn ; Autophagy ; Beclin-1 ; physiology ; Fibroblasts ; Glucose ; Microtubule-Associated Proteins ; Mitochondrial Proteins ; Rats ; Rats, Sprague-Dawley

In order to successfully express the carbonyl reductase gene mldh in Bacillus subtilis and complete coenzyme regeneration by B. subtilis glucose dehydrogenase, the promoter PrpsD and the terminator TrpsD from B. subtilis rpsD gene were used as the expression cassette to be a recombinant plasmid pHY300plk-PrpsD-TrpsD. After that, the carbonyl reductase gene mldh was inserted into the previous plasmid and a plasmid pHY300plk-PrpsD-mldh-TrpsD was achieved, followed by transformed into B. subtilis Wb600 to obtain a recombinant B. subtilis Wb600 (pHY300plk-PrpsD-mldh-TrpsD). Subsequently, the results for whole-cell biotransformation from recombinant B. subtilis showed that it could be used to catalyze MAK (1-phenyl- 1-keto-2-methylaminopropane) to d-pseudoephedrine in the presence of glucose. The yield of d-pseudoephedrine could be up to 97.5 mg/L and the conversion rate of MAK was 24.1%. This study indicates the possibility of biotransformation production of d-pseudoephedrine from recombinant B. subtilis.
Alcohol Oxidoreductases ; genetics ; Bacillus subtilis ; genetics ; metabolism ; Glucose 1-Dehydrogenase ; chemistry ; metabolism ; Mutagenesis, Insertional ; Pseudoephedrine ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Recombination, Genetic

BACKGROUND: Insulin resistance is an integral feature of metabolic syndromes, including obesity, hyperglycemia, and hyperlipidemia. In this study, we evaluated whether the aloe component could reduce obesity-induced inflammation and the occurrence of metabolic disorders such as blood glucose and insulin resistance. METHODS: Male C57BL/6 obese mice fed a high-fat diet for 54 days received a supplement of aloe formula (PAG, ALS, Aloe QDM, and Aloe QDM complex) or pioglitazone (PGZ) and were compared with unsupplemented controls (high-fat diet; HFD) or mice fed a regular diet (RD). RT-PCR and western blot analysis were used to quantify the expression of obesity-induced inflammation. RESULTS: Aloe QDM lowered fasting blood glucose and plasma insulin compared with HFD. Obesity-induced inflammatory cytokine (IL-1beta, -6, -12, TNF-alpha) and chemokine (CX3CL1, CCL5) mRNA and protein were decreased markedly, as was macrophage infiltration and hepatic triglycerides by Aloe QDM. At the same time, Aloe QDM decreased the mRNA and protein of PPARgamma/LXRalpha and 11beta-HSD1 both in the liver and WAT. CONCLUSION: Dietary aloe formula reduces obesity-induced glucose tolerance not only by suppressing inflammatory responses but also by inducing anti-inflammatory cytokines in the WAT and liver, both of which are important peripheral tissues affecting insulin resistance. The effect of Aloe QDM complex in the WAT and liver are related to its dual action on PPARgamma and 11beta-HSD1 expression and its use as a nutritional intervention against T2D and obesity-related inflammation is suggested.
11-beta-Hydroxysteroid Dehydrogenase Type 1 ; Aloe ; Animals ; Blood Glucose ; Blotting, Western ; Cytokines ; Diabetes Mellitus, Type 2 ; Diet ; Diet, High-Fat ; Fasting ; Glucose ; Humans ; Hyperglycemia ; Hyperlipidemias ; Inflammation ; Insulin ; Insulin Resistance ; Liver ; Macrophages ; Male ; Mice ; Mice, Obese ; Obesity ; Plasma ; PPAR gamma ; RNA, Messenger ; Thiazolidinediones ; Triglycerides

OBJECTIVETo explore the effects of glucose concentration fluctuation on function of cultured bovine arterial endothelial cells and underlying mechanism.

METHODSThe thoracic aorta of newborn calf was used for primary endothelial cells culture. Cells were divided into 3 groups and cultured for 48 h: control group (C, 5.5 mmol/L), constant high glucose group (HG, 30 mmol/L) and glucose fluctuation (GF, three circles of 2 h 30 mmol/L followed by 3 h 5.5 mmol/L, 30 mmol/L overnight, repeat the whole procedure on the following day) groups. The membranes fluidity of endothelial cells was detected by fluorescence polarization method. The contents of sorbierite, aldose reductase (AR), sorbitol dehydrogenase (SDH) and advanced glycation end products (AGEs) were measured. RAGE, eNOS and ET-1 mRNA expressions were detected by semi-quantitative RT-PCR.

RESULTSThe membranes fluidity of endothelial cells in HG or GF group were significantly decreased compared with the control group (all P < 0.01) and significantly lower in GF group than those in HG group (all P < 0.01). Sorbierite, AR and AGEs concentrations were significantly higher in HG and GF groups than those in control group (all P < 0.01) and AR and AGEs concentrations were significantly higher in GF group than that in HG group (all P < 0.01). SDH of endothelial cells in HG or GF group were decreased compared with the control group and lower in GF group than in HG group (all P < 0.05). In addition, the mRNA levels of RAGE, eNOS and ET-1 were significantly upregulated compared with the control group (all P < 0.01).

CONCLUSIONSGlucose concentration fluctuation can result in more severe bovine arterial endothelial cells dysfunction than high glucose via activating polyols metabolic pathways, upregulating the expression of AGEs, eNOS and ET-1. Therefore, glucose concentration fluctuation might play a crucial role on macrovascular complications of diabetes.

Aldehyde Reductase ; analysis ; Animals ; Aorta, Thoracic ; cytology ; Cattle ; Cells, Cultured ; Endothelial Cells ; metabolism ; pathology ; Endothelin-1 ; analysis ; Endothelium, Vascular ; cytology ; metabolism ; Glucose ; metabolism ; Glycation End Products, Advanced ; analysis ; L-Iditol 2-Dehydrogenase ; analysis ; Membrane Fluidity ; Nitric Oxide Synthase Type III ; analysis

11-beta-Hydroxysteroid Dehydrogenase Type 1 ; genetics ; metabolism ; Adipose Tissue, White ; drug effects ; Animals ; Animals, Newborn ; Blood Glucose ; metabolism ; Body Weight ; drug effects ; Drug Synergism ; Eating ; drug effects ; Fenofibrate ; pharmacology ; Hypoglycemic Agents ; pharmacology ; Hypolipidemic Agents ; pharmacology ; Insulin Resistance ; Lipids ; blood ; Lipoprotein Lipase ; genetics ; metabolism ; Liver ; drug effects ; Metabolic Syndrome ; chemically induced ; metabolism ; pathology ; Mice ; Mice, Inbred ICR ; RNA, Messenger ; metabolism ; Sodium Glutamate ; Thiazolidinediones ; pharmacology