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BackgroundThe connection between gut microbiota and metabolism and its role in the pathogenesis of diabetes are increasingly recognized. The objective of this study was to quantitatively measure Bifidobacterium and Lactobacillus species, members of commensal bacteria found in human gut, in type 2 diabetic patients (T2D) patients from Southern China.MethodsFifty patients with T2D and thirty control individuals of similar body mass index (BMI) were recruited from Southern China. T2D and control subjects were confirmed with both oral glucose tolerance test (OGTT) and HbA(1c) measurements. Bifidobacterium and Lactobacillus species in feces were measured by real-time quantitative PCR. Data were analyzed with STATA 11.0 statistical software.ResultsIn comparison to control subjects T2D patients had significantly more total Lactobacillus (+18%), L. bugaricus (+13%), L. rhamnosum (+37%) and L. acidophilus (+48%) (P < 0.05). In contrast, T2D patients had less amounts of total Bifidobacteria (-7%) and B. adolescentis (-12%) (P < 0.05). Cluster analysis showed that gut microbiota pattern of T2D patients is characterized by greater numbers of L. rhamnosus and L. acidophillus, together with lesser numbers of B. adolescentis (P < 0.05).ConclusionThe gut microflora in T2D patients is characterized by greater numbers of Lactobacillus and lesser numbers of Bifidobacterium species.

BackgroundThe connection between gut microbiota and metabolism and its role in the pathogenesis of diabetes are increasingly recognized. The objective of this study was to quantitatively measure Bifidobacterium and Lactobacillus species, members of commensal bacteria found in human gut, in type 2 diabetic patients (T2D) patients from Southern China.MethodsFifty patients with T2D and thirty control individuals of similar body mass index (BMI) were recruited from Southern China. T2D and control subjects were confirmed with both oral glucose tolerance test (OGTT) and HbA(1c) measurements. Bifidobacterium and Lactobacillus species in feces were measured by real-time quantitative PCR. Data were analyzed with STATA 11.0 statistical software.ResultsIn comparison to control subjects T2D patients had significantly more total Lactobacillus (+18%), L. bugaricus (+13%), L. rhamnosum (+37%) and L. acidophilus (+48%) (P < 0.05). In contrast, T2D patients had less amounts of total Bifidobacteria (-7%) and B. adolescentis (-12%) (P < 0.05). Cluster analysis showed that gut microbiota pattern of T2D patients is characterized by greater numbers of L. rhamnosus and L. acidophillus, together with lesser numbers of B. adolescentis (P < 0.05).ConclusionThe gut microflora in T2D patients is characterized by greater numbers of Lactobacillus and lesser numbers of Bifidobacterium species.

This study is conducted to explore new methods to perform surface biomodification of titanium implants and improve osteogenic efficiency. Methods An RGD peptide and chitosan (CS) were combined by acylation reaction, forming RGD-CS. An RGD-CS/pDNA complex was subsequently prepared using a complex coacervation method and grafted on a pure titanium surface after physical and biochemical treatments were performed. The chemical structural characteristics of RGD-CS were evaluated using an infrared spectrometer and an elemental analyzer. The shape of this complex was then assessed by gel electrophoresis combined with atomic force microscopy. The grafting effect of this complex on the titanium surface was detected by EB staining. Results CS and RGD peptides were coupled by an amide bond. The RGD-CS/pDNA complex was completely composited at N /P⩾2. Atomic force microscopy results showed that the morphology of this complex was mainly spherical. EB staining experiments showed that this complex was successfully grafted on the titanium plate. Conclusion RGD peptide-modified CS can be used as a titanium implant surface plasmid package carrier of pDNA. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the cutoff value of HbA1c for predicting diabetes and prediabetes in a Chinese high risk population aged over 45.Methods: A total of 619 people aged over 45 without diabetes were randomly recruited to complete Finnish Diabetes Risk Score (FINDRISC) questionnaire. 208 high-risk individuals (defined by Diabetes Risk Score >=9) had OGTT and HbA1c determined at the same time.Results: In a Chinese population aged over 45, the best cutoff value of HbA1c for detecting diabetes and prediabetes was 5.8% and 5.4% respectively. The area under the receiver operating characteristic (AUROC) curve of HbA1c for detecting diabetes was 0.85 (95% CI: 0.80-0.90) and prediabetes was 0.62 (95% CI: 0.54-0.70). The combined use of HbA1c and fasting blood glucose (FPG) had larger AUROC than HbA1c alone (0.88, 95%CI: 0.83-0.92 in detecting diabetes vs 0.75, 95% CI: 0.67-0.82 in prediabetes), and had a higher sensitivity in predicting diabetes and higher specificity and positive predictive value (PPV) in predicting prediabetes. However, the AUROC between HbA1c alone and combined use in predicting diabetes was not significantly different (p=0.173).Conclusions: FINDRISC is feasible tool to screen people who are at high risk of diabetes. The cutoff values of HbA1c to diagnose diabetes and prediabetes in a Chinese high risk population aged over 45 were 5.8% and 5.4%, respectively. The sensitivity and specificity of HbA1c for detecting diabetes and prediabetes was relatively low, so that the combined use of HbA1c and FPG may be more effective in prediction. [ABSTRACT FROM AUTHOR]

Background: Type 2 diabetes mellitus (T2DM)-associated mortality and morbidity are strongly dependent on glycemic control. With T2DM prevalence increasing in China, we aimed to assess glycemic control rates in Chinese T2DM outpatients., Material/methods: A total of 9065 adult T2DM outpatients (5035 men) were assessed in 26 Chinese medical centers between August 2010 and April 2012. Patients were stratified according to BMI (kg/m2): <24, 24-28, and >28. Successful glycemic control was defined as glycated hemoglobin A1c (HbA1c) ≤7% or fasting plasma glucose (FPG) <7.0 mmol/L., Results: Among the participants included in this study, 2939 had BMI <24, 3361 had BMI of 24-28, and 2764 had BMI >28. The glycemic control rate was only 32.6%, and the triple control rate for glycemia, blood pressure, and lipidemia was only 11.2%. Glycemic control rates by BMI group were 33.7% (<24), 33.8% (24-28), and 30.2% (>28) (p=0.005), and corresponding incidences of cardiovascular diseases (CVD) were 12.2%, 15.7%, and 15.9% (p<0.001). Multivariate logistic regression analysis demonstrated that older age (p<0.001), higher BMI (p=0.026), larger waist circumference (p<0.001), less education (p<0.001), and recent diagnosis (p<0.001) were independent risk factors for poor glycemic control., Conclusions: The T2DM glycemic control rate in China is currently low, especially in older obese patients with poor education and recent diagnosis.

Objective: This study is conducted to explore new methods to perform surface biomodification of titanium implants and improve osteogenic efficiency., Methods: An RGD peptide and chitosan (CS) were combined by acylation reaction, forming RGD-CS. An RGD-CS/pDNA complex was subsequently prepared using a complex coacervation method and grafted on a pure titanium surface after physical and biochemical treatments were performed. The chemical structural characteristics of RGD-CS were evaluated using an infrared spectrometer and an elemental analyzer. The shape of this complex was then assessed by gel electrophoresis combined with atomic force microscopy. The grafting effect of this complex on the titanium surface was detected by EB staining., Results: CS and RGD peptides were coupled by an amide bond. The RGD-CS/pDNA complex was completely composited at N/P > or = 2. Atomic force microscopy results showed that the morphology of this complex was mainly spherical. EB staining experiments showed that this complex was successfully grafted on the titanium plate., Conclusion: RGD peptide-modified CS can be used as a titanium implant surface plasmid package carrier of pDNA.

Objective: To investigate the effect of cetylpyridinium chloride buccal tablets on halitosis induced by oral conditions., Methods: With Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum as the testing bacteria, the minimal inhibitory concentration (MIC) of cetylpyridinium chloride buccal tablets was determined using minute amount serial dilution test. The production of volatile sulfur compounds (VSCs) was measured using sulfide detector halimeter in the anaerobic bacteria culture at 4 and 8 h after addition of the tablets. The effect of the tablets in suppressing odor production by mouth-borne halitosis bacteria was assessed using cysteine challenge test in healthy volunteers, and the effectiveness was evaluated by measuring the reduction in VSCs production and the duration of the effect., Results: Cetylpyridinium chloride buccal tablets inhibited the growth of all the 3 bacteria. The tablets obviously inhibited VSCs production by the 3 bacteria with a effect similar to chlorhexidine. Compared with distilled water gargle, the buccal tablets significantly reduced cysteine-induced VSCs production level in the healthy volunteers (P<0.05), and the effect lasted for 230 min., Conclusion: Cetylpyridinium chloride tablets can obviously suppress bacteria responsible for oral halitosis and produce good effects in the treatment of halitosis induced by oral conditions.

Objective: To investigate the role of Toll-like receptor 4 (TLR4) signaling pathway in the activation of rennin- angiotensin system (RAS) in adipose cells., Methods: 3T3-L1 cells induced with isobutylmethylxanthine, insulin and dexamethasone to differentiate into adipocytes were stimulated by LPS with or without irbesartan pretreatment. The expression levels of TLR4, angiotensinogen (AGT) and angiotensin II receptor type 1 (ATlR) mRNA in 3T3-L1 cells was determined by RT-PCR, and their protein expressions were detected with Western-blotting. Immunofluorescence double staining was used to observe the translocation of NF-κB p65 subunit in the cells., Results: Stimulation with LPS dose- and time-dependently increased the mRNA and protein expressions of TLR4, AGT and AT1R. LPS exposure resulted in enhanced translocation of NF-κB p65 subunit in the adipose cells, which was attenuated by irbesartan pretreatment., Conclusion: Activation of TLR4 signaling pathway may trigger the activation of local RAS in adipose cells.

Objective: To study the correlation between adipocytokines levels and metabolic syndrome (MS) in patients with newly diagnosed type 2 diabetes mellitus (T2DM)., Methods: Sixty-eight patients with newly diagnosed T2DM, including 51 cases with MS and 17 without MS, were examined for blood pressure (BP), fasting blood glucose (FBG), fasting insulin (FINS), insulin resistance index (IRI), waist to hip ratio (W/H), body mass index (BMI), and serum adipocytokine levels (IL-6, vaspin, and adiponectin)., Results: The diabetic patients with MS had higher BMI, HbA1c, FBG, FINS, IRI, TG, TC, and SBP than those without MS. Serum IL-6 level was higher but adiponectin level was lower in patients with MS than in those without MS. There was no significant difference in vaspin level between the two groups. Adiponectin level was positively correlated with TG (r=-0.30, P=0.02) and inversely with BMI (r=-0.47, P=0.39) and HOMA-IR (r=-0.30, P=0.03); vaspin level was positively correlated with HOMA-IR (r=0.347, P=0.02) and inversely with HDL-L (r=-0.45, P=0.01); IL-6 level was positively correlated with LDL-L (r=0.18, P=0.25) and inversely with HDL-L (r=-0.45, P=0.01)., Conclusion: Adiponectin and IL-6 levels are closely related to MS, but the relationship between vaspin and MS needs further investigation.

Objective: To study the effect of pretreatment with apoptotic donor spleen cells on spleen lymphocyte function of recipient rats undergoing islet transplantation to explore new approaches to prolong islet graft survival., Methods: Apoptotic spleen cells from donor rats were obtained by exposure to γ-ray irradiation from (60)Co. Diabetic SD rat models were randomly divided into 4 groups to receive tail vein injections with saline (group A), normal cells (group B), apoptotic donor cells (group C), or necrotic donor cells (group D). One week later, orthotopic transplantation of islets under the renal capsule was performed. Before and at 1 and 2 weeks after islet transplantation, the recipient rats were examined for proliferative activity of spleen lymphocytes with CFSE cell staining and for IL-2 and IL-10 expressions in the cells using ELISA., Results: Pretreatment with donor apoptotic cells significantly suppressed the proliferative activity of recipient spleen lymphocytes before and at 1 and 2 weeks after islet transplantation as compared with the other three groups (P<0.05). The level of IL-2 was significantly decreased while IL-10 increased in apoptotic donor cell pretreatment group compared with those in the other 3 groups at each time point of observation., Conclusion: The effect of pretreatment with apoptotic donor cells on recipient spleen lymphocytes suggest an important role of apoptotic donor spleen cells in immune tolerance of grafts.

Objective: To study the signaling pathways associated with lipopolysaccharide (LPS)-induced inflammation in islet micro-endothelial cells (IMECs) and the mechanism of pravastatin intervention., Methods: IMECs exposed to LPS, SB203580, pravastatin, or SB203580+pravastatin were examined for cell apoptosis with Hoechst staining and flow cytometry and for expression levels of total-p38, photophosphorylation-p38 (p-p38) and iNOS with Western blotting., Results: The apoptosis rate and expression levels of total-p38, p-p38, iNOS in IMECs all increased after LPS exposure. Pravastatin, SB203580, and their combination significantly attenuated LPS-induced enhancement of cell apoptosis and total-p38, p-p38, and iNOS expressions in IMECs., Conclusion: LPS-induced inflammatory toxicity in IMECs is associated with the activation of P38MAPK and iNOS/NO signaling pathways. Pravastatin can inhibit these pathways and suppress the apoptosis and necrosis of IMECs to relieve the cell inflammatory injuries.

Objective: To investigate the effect of advanced oxidation protein products (AOPP) on proliferation, differentiation, and oxidative stress in rat osteoblasts., Methods: The cell proliferation and differentiation of osteoblasts isolated from neonatal SD rat skull were evaluated following treatment with different concentrations (50, 100, 200, and 400 µg/ml) of AOPP using CCK-8 kit and ALP assay kit, respectively. The levels of reactive oxygen species (ROS) in the treated cells were analyzed using 2', 7'-dichlorofluorescin diacetate, and the transcription levels of ALP, collagen I and RAGE were assessed using real-time PCR., Results: Compared with the control group, AOPP-treated osteoblasts showed obviously inhibited proliferation and differentiation with down-regulated expressions of ALP and collagen I and increased ROS production and RAGE expression., Conclusion: AOPP can inhibit the proliferation and differentiation of rat osteoblasts partially by up-regulating RAGE and inducing ROS production.

Objective: To investigate the role of TRB3 in the inhibitory effect of fenofibrate against the proliferation of glomerular mesangial cell induced by high glucose., Methods: Rat glomerular mesangial cells (MCs) were cultured in the presence of 5.5 mmol/L glucose (normal control), 25 mmol/L glucose (high glucose group), or high glucose along with 10, 50, or 100 µmol/L fenofibrate. Cell counting kit-8 (CCK-8) assay was used to evaluate cell proliferation, and Hoechst 33258 staining was employed to determine chromatin distribution in the MCs. Flow cytometry was performed to analyze the cell cycle changes in different groups. The expressions of TRB3 and P-AKT in different groups were detected using immunocytochemistry and Western blotting., Results: High glucose induced obvious proliferation of the MCs (P<0.001), which was significantly inhibited by fenofibrate in a concentration-dependent manner (P<0.001). The MCs exposed to fenofibrate presented with typical apoptotic morphologies and cell cycle arrest at G1/S phase. Low levels of TRB3 expression was detected in the normal control and high glucose groups, whereas in the 3 fenofibrate groups, TRB3 expression increased and P-AKT expression decreased as fenofibrate concentration increased., Conclusion: Fenofibrate can promote TRB3 expression in rat MCs. TRB3 causes cell cycle arrest at G1/S phase by inhibiting AKT phosphorylation to result in suppressed proliferation of the MCs.

Objective: To investigate the effect of liraglutide on the inflammatory cytokine interleukin-1β (IL-1β) and apoptotic factor caspase-3 expression in the pancreatic islets of OLETF rats with impaired glucose tolerance., Methods: Twelve-week-old OLETF rats were randomized into 4 groups and received intraperitoneal injections of saline or liraglutide at 50, 100, or 200 µg/kg twice daily for 12 weeks. Eight LETO rats served as the normal control group and received saline injection. After the treatments, the rats were examined for fasting and 30 min plasma insulin during OGTT test, and the expression levels of IL-1β and caspase-3 mRNA and protein in the pancreatic islets were detected by real-time PCR and Western blotting, respectively., Results: Compared with the saline group, liraglutide significantly decreased the expressions of IL-1β and caspase-3 mRNA and protein, and significantly improved the blood glucose, islet β function and early-phase insulin secretion index in OLETF rats., Conclusions: Liraglutide can improve islet function and glucose metabolism partially by inhibiting islet IL-1β expression to delay or prevent the development of diabetes in OLETF rats.

Objective: To determine the changes in fecal Bifidobacterium species in patients with type 2 diabetes in comparison with healthy individuals., Methods: The bacterial DNA were extracted from the fecal samples from 50 type 2 diabetic patients and 30 healthy individuals. Real-time quantitative PCR was employed to determine the copy numbers of the bacteria in the fecal samples using 16S rRNA-targeted genus- and species-specific PCR primers for a selected group of fecal Bifidobacterium species including total Bifidobacterium, B.longum, B.breve, B.adolescent, and B. infantis., Results: The diabetic group had significantly lower copy numbers of total Bifidobacterium and B.adolescent compared to the healthy individuals (P<0.05)., Conclusion: Type 2 diabetic patients have a lowered number of Bifidobacterium species in the gut microflora.

Objective: [corrected] To characterize the insulinotropic action of hippocampal cholinergic neurostimulating peptide (HCNP) and analyze the role of type 3 muscarinic receptor (M(3)R) pathway in the action of HCNP., Methods: INS-1 cells were incubated in routine RPMI 1640 medium (control group), RPMI 1640 supplemented with 50 pg/ml synthetic HCNP (HCNP group), or HCNP-containing medium with the addition of PMA 18 h prior to insulin release assay. The insulin levels in the medium was measured using radioimmunoassay following stimulation with different concentrations of glucose. Real-time quantitative PCR was used for detecting the gene expression of HCNP-pp, choline acetyltransferase (ChAT) and M(3)R in HCNP group and control group., Results: After stimulation with different concentrations of glucose (5.6 and 16.7 mmol/L), HCNP group showed significantly higher insulin levels than the control and HCNP+ PMA groups. Compared with those in the control group, the mRNA levels of HCNP-pp, ChAT, and M(3)R were all lowered in HCNP group., Conclusion: HCNP can promote insulin release in INS-1 cells by increasing ChAT activity and activating M(3)R, and this effect is inhibited by PMA.