Objective To determine the effects of glucose concentration on G 6PD activity and respiratory burst of normal hu-man′s neutrophils in vitro .Methods Normal human′s neutrophils were cultured in different glucose concentration for 8 hours ,as-sayed G6PD activity by spectrophotometric method and determining ROS content by fluorescent probe DCFH-DA .Results G6PD activity and ROS of 15 mmol/L group and 25 mmol/L group were significant lower than before ,when the 5 mmol/L group and L-GLU group didn′t have significant change with time goes by .And G6PD activity and ROS of 25 mmol/L group were the lowest in all groups(P<0 .01) .Conclusion High glucose may induce G6PD activity decreased and cause respiratory burst dysfunction as a stimulating factor .The stimulation intensity was increased with the increase of glucose concentration .It′s the probable mechanism on susceptibility to infections in patients with diabetes mediated by dysfunction of respiratory burst in leucocyte .

OBJECTIVETo observe the changes in glucose 6-phosphate dehydrogenase (G6PD) activity, cAMP and respiratory burst function in THP-1 cells exposed to high glucose and identify the possible signaling pathways to mediate these changes.

METHODSTHP-1 cells were treated with high glucose, high glucose plus the PKA inhibitor (PKI), or normal glucose plus Forskolin. The changes in the G6PD activity and cAMP in the exposed cells were assayed using the spectrophotometric method, and the reactive oxygen species (ROS) content in the cell culture was determined using the fluorescent probe DCFH-DA. Western blotting was employed to examine the expression of phosphorylated p47(phox) in the cells.

RESULTSCompared with the normal control cells, the cells exposed to high glucose and to normal glucose and Forskolin showed a significantly lowered G6PD activity, ROS content and expression of phosphorylated p47(phox), but with a increased cAMP content (P<0.01). High glucose exposure in the presence of PKI caused no significant changes in G6PD activity, ROS level, phosphorylated p47(phox) or cAMP compared to those in the normal control cells (P>0.01).

CONCLUSIONHigh glucose causes inhibition of G6PD activity in THP-1 cells via activation of PKA and thus leads to respiratory burst dysfunction, which is the probable mechanism underlying the lowered leucocyte function and susceptibility to infections in diabetic patients.

Cell Line ; Cyclic AMP ; metabolism ; Cyclic AMP-Dependent Protein Kinases ; antagonists & inhibitors ; metabolism ; Glucose ; adverse effects ; Glucosephosphate Dehydrogenase ; metabolism ; Humans ; Phosphorylation ; Respiratory Burst ; drug effects ; Signal Transduction ; drug effects

Objective To clone the human Islet neogenesis associated protein(rhINGAP)gene,express the gene extraeellulary in Pichia yeast for.further study on biological function and animal test on INGAP.Methods INGAP gene Was amplified with PCR and inserted into the recombinant plasmidα/pUC18.Then,the fusion gene of α and INGAP was digested and inserted into the expression plasmid pPIC9K.The positive recombinant plasmid which integrated INGAP Was confirmed by restriction enzyme digestion and sequencing,and it Was linearized with Sal Ⅰ digestion and transfered into the yeast host strain GS115 through electroporation.The yeast transformants that harbor the desired gene INGAP with high copy were selected by the auxotroph mediam G418,and verified by PCR.The condition of hake-flask culture was optimized,and the recombinant human INGAP Was induced expression with methanol as the only Carbone source.The antigen activity of the desired protein Was detected by Western blotting and ELISA method.Results Recombinant plasmid αINGAP/pPIC9K were successfully constructed and three positive Pichia yeast transformants were obtained.The expressed protein had satisfactory antigen activity,which Was confirmed by the Western blotting and ELISA method.Conclusions Pichia yeast expressing human Islet neogenesis associated protein (rhINGAP)gene was successfully constructed.

ObjectiveTo explore the mechanism of estrogen inhibiting osteoblast apoptosis induced with serum hungry.MethodsOsteoblasts of the second or third generation from newly born SD rats calvaria were divided randomly into three groups: control group,serum hungry group,serum hungry with estrogen group.Cells of each group were incubated for 1,2,3,5,7 or 14 d,and then were stained immunohistochemically.The rates of positive cells of each group were analyzed.ResultsThere was a little positive expression of Bax,Bcl2 and Fas in control group.The expression of Bax and Fas were significantly increased(P<0.05)in serum hungry group,peak time was 14 d,but the expression of Bcl-2 were not affected.Compared with that of serum hungry group,the expression of Bax and Fas significantly decreased(P<0.05) in serum hungry and estrogen group,peak time was still 14 d,while that of Bcl-2 increased(P<0.05).ConclusionSerum hungry can increase the expression of Bax and Fas in osteoblast,that can be inhibited by estrogen.Estrogen can also increase the expression of Bcl-2 in osteoblast.All of these may play a role in inhibiting osteoblast apoptosis induced with serum hungry.

OBJECTIVETo investigate the effect of blood glucose instability on respiratory burst of leukocytes in patients with type 2 diabetes (T2DM).

METHODSForty-five patients with T2DM were divided into 3 groups after continuous glucose monitoring for 72 h with glucose wavy coefficient <1.5 (n=11), between 1.5 and 3.0 (n=19), and >3.0 (n=15). Peripheral blood neutrophils were isolated from the diabetic patients and normal control subjects for assay of glucose 6-phosphate dehydrogenase (G6PD) with a spectrophotometric method, detecting G6PD mRNA expression by real-time PCR, and determining reactive oxygen species level using the fluorescent probe DCFH-DA.

RESULTSCompared with the normal control group, the diabetic patients showed significantly lowered G6PD activity (F=78.739, P<0.05) and ROS level (F=384.962, P<0.05) but significantly increased G6PD mRNA expression (F=269.612, P<0.01). These changes were significantly correlated with the blood glucose wavy coefficients.

CONCLUSIONThe fluctuation of blood glucose in T2DM patients can decrease G6PD activity and lead to functional decline of the respiratory burst.

Blood Glucose ; chemistry ; Case-Control Studies ; Diabetes Mellitus, Type 2 ; metabolism ; Glucosephosphate Dehydrogenase ; metabolism ; Humans ; Neutrophils ; metabolism ; Pentose Phosphate Pathway ; Reactive Oxygen Species ; metabolism ; Respiratory Burst

[Objective]To discuss the intension and extension of five excesses & five deficiency in Canon of Medicine, and to investigate its guiding value in clinical practice.[Method]By consulting document, and to know how these ancient Chinese medicine doctors to understand and analyse five excesses & five deficiency in Canon of Medicine, with deep demonstration, and finally to prove by clinical use.[Result] Most ancient Chinese medicine doctors agreed that five excesses & five deficiency was clinical critical disease, usually signified an ominous prognosis, and finally led to die.On the one hand it was excessive pathogenic qi rampant, disturbing five organs; on the other hand it was asthenia of health qi, and fall of five organs.In summary, the pathogenesis of five excesses was five organs being attacked, pathogenic qi impatented, excessive heat, riddlling in both internal and external, main pathogenesis was internal stagnancy of pathogenic heat, leading exhaustion of yin or yang, while the pathogenesis of five deficiency was real yang exhausted, real yin exhausted. Therefore treatment principle of five excesses & five deficiency has guiding significance for clinician, especially to protect stomach-qi in clinical practice.[Conclusion]The Canon of Medicine first proposed the theory of five excesses & five deficiency, including the conception, prognosis, and treatment principle. Five excesses & five deficiency were the earliest classification of excesses & deficiency of five-viscera, and it established the foundation of excesses & deficiency syndrome differentiation in TCM, and It had guiding significance to identify, judge and treatment of TCM clinical critical disease, and it needed paying more attention and further research.

Objectives To investigate the epidemiological and clinical characteristics of dyslipidemia as well as its treatment and influence on accompanying diseases in impaired glucose status among inpatients. Methods A cross-sectional survey was conducted among the inpatients registered in ten university hospitals of Guangdong, China during the week before the Diabetes Day in 2004. The fasting blood glucose (FBG), lipid profiles, BMI, waist to hip ratio (WHR) and concomitant disorders of the first screen during the hospitalization period were recorded. Those who had FBG level from 5.6 to 6. 9 mmol/L and not been previously diagnosed diabetes (PDM) underwent oral glucose tolerance test (OGTF). Results Of the 8753 inpatients investigated, 1067 eases had complete medical records(CMR case) including PDM cases and previously non-diagnosed diabetes ones with FBG ≥ 5. 6 mmol/L. Of the previously non-diagnosed diabetes cases with FBG levels from 5.6 to 6.9 mmmol/L, 65.8% accepted OGTT. Of the CMR cases, 41.9% had PDM, 21.7% was newly diagnosed diabetes mellitus (NDM), 29. 1% had impaired glucose regulation (IGR) and only 7.3% had normal glucose tolerance (NGT). The TG levels in NDM and PDM group were higher than those in IGR and NGT group (P < 0.05, respectively). The HDL-C levels in IGR, NDM and PDM group were lower than those in NGT group (P < 0.05, respectively). Sixty-nine point six percent of the diabetes mellitus (DM) inpatients was accompanied with dyslipidemia and the rate was higher than those in NGT (56.4%) and IGR inpatients (52.5%, P <0.05, respectively). Only 22. 8% of the PDM inpatients underwent treatment of dyslipidaemia and just 3.4% achieved the target suggested by the guideline of ATP-Ⅲ. BMI was higher and waistline longer in the PDM and NDM inpatients than those in the NGT cases (P <0.05, respectively). Seventy-two point eight percent of the PDM inpatients was complicated with more than one type of vascular diseases. Nine point seven percent and 0. 2% of the NDM inpatients were tormented by diabetic nephropathy and diabetic retinopathy respectively. Conclusions More inpatients with accompany DM or IGR had concomitant dyslipidemia than those with NGT, which included hypertriglyccridemia, hypo-high-density lipoproteinemia and metabolic syndrome. Concomitant vascular diseases were more frequently found in PDM inpatients than in the others. Some of the NDM and IGT inpatients were complicated with microvascular diseases.

Impaired eady phase insulin secretion is an important reason for leading to postprandial hyperglycemia.Nateglinide is a rapid-acting insulin secretagogue,which reduces postprandial blood glucose of type 2diabetic patient by restoring early phase insulin secretion.The efficacy and safety have been fully verified by clinical administration and it is more widely used to treat type 2 diabetic patients.Both sulfonylureas and glinides were named insulin secretagogue agents and regarded as alternative first-line drugs in the 2010 Chinese Guideline for treatment of type 2 diabetes.AACE/ACE Consensus statement claimed that glinides would be one of the important choices after metformin.In order to further guide the clinical application of nateglinide,16 national specialists in the field of endocrinology and metabolism of China discussed,drafted,and edited this consensus.The current consensus combined clinical evidences at home and abroad.systematically reviewed and summarized tlle results of these studies about nateglinide.It will provide guiding recommendations and reference concerning how to reasonably and effectively use nateglinide in the clinical practice.

OBJECTIVE: To investigate the effect of glucagon-like peptide 1 receptor agonists (GLP-1RAs) on body fat redistribution and muscle mass in overweight/obese patients with type 2 diabetes (T2DM). METHODS: We retrospectively analyzed the data of 76 patients with body mass indexes (BMI)≥24 kg/m, who had an established diagnosis of T2DM in our department between December, 2014 and September, 2015. We divided these patients according to their BMI in overweight group (BMI of 24-27.9 kg/m, =14), obese group (BMI of 28-31.9 kg/m, =35) and severely obese group (BMI≥32 kg/m, =27). All the patients received treatment with GLP-1RAs (Exenatide or Liraglutide) for 3.0 to 29.0 weeks (mean 8.9 weeks), and their blood glucose, HbA1c and serum lipids were analyzed. For each patient, the fat and muscle masses were analyzed using a human body composition analyzer (JAWON-IOI353, Korea) before and after GLP-1RAs treatment. RESULTS: Treatment with GLP-1RAs significantly decreased BMI and visceral adiposity index (VAI) in all the patients in the 3 groups ( < 0.05). The treatment significantly decreased the body weight in the overweight group and obese group by 2.70 kg (0.60-4.95 kg) and 2.65 kg (1.45-6.40 kg), respectively ( < 0.05), and significantly decreased the waist-to-hip ratio (WHR) in the overweight group ( < 0.05). The obese and severely obese patients showed significantly decreased percentage body fat (including both subcutaneous and visceral fat) and increased muscle mass after the treatment ( < 0.05). Compared with those in the overweight group, the percentage body fat and VAI were significantly decreased in the obese group after the treatment ( < 0.05), and the percentage of subcutaneous fat reduced and the muscle ratio increased more obviously in the obese and severely obese patients ( < 0.05). CONCLUSIONS GLP-1RAs treatment can significantly lower BMI and improve body fat distribution in obese patients with T2DM, especially in patients with a greater BMI.
Adipose Tissue ; Body Mass Index ; Diabetes Mellitus, Type 2 ; Glucagon-Like Peptide-1 Receptor ; Humans ; Hypoglycemic Agents ; Obesity ; Overweight ; Retrospective Studies