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Objective To observe the effect of exogenous insulin on the expression of P-glycoprotein (P-gp)and the secretion of insulin in pancreatic beta cells (INS-1 832/13).Methods Insulinoma cells (INS-1 832/13) were cultured with 0.5 μmol/L exogenous insulin for 30 days.MTT assay was used to measure cell viability.Quantitative RT-PCR and western blot were used to detect the expression of P-gp mRNA and protein respectively,and glucose stimulated insulin secretion (GSIS) were measured by radioimmunoassay.Results Compared with control group,0.5 μmol/L exogenous insulin promoted the viability of INS-1 832/13 cells [(102.00±12.99) vs (356.00±35.51),P<0.05] and accelerated P-gp expression [(107.50±17.08) vs (307.50±44.25)] both at mRNA and protein levels [(105.00±12.91) vs (192.50±35.94),P<0.05].Glucose stimulated insulin secretion was positively correlated with P-gp expression level,but had no significant effect on basal insulin secretion.Conclusion Exogenous insulin can promote the secretion function of INS-1 832/13 cells,and the mechanism may be related to the expression of P-gp.
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Objective To explore the effect of Shenling Baizhu power on inflammatory factors and immune function for recurrent aphthous ulcer. Methods Ninety patients with recurrent aphthous ulcer in our hospital were divided into control group and study group according to random number table method, with 45 cases each group.The control group were given oral vitamin B2, levamisole and cetyl pyridinium chloride gargle solution therapy, and the study group were given Shenling Baizhu power on the basis of control group.After one week treatment, the clinical curative effect were observed, the serum inflammatory factors levels of tumor necrosis factor alpha ( TNF-α) , interleukin 2 ( IL-2 ) and interleukin 6 ( IL-6 ) were determined by Elisa method, the levels of peripheral blood T cell subsets CD3 +, CD4 +, CD8 +and natural killer cells( NKT) were determined by flow cytometry instrument, and the recurrence rate were observed after one year follow-up.Results After one week treatment, the total effective rate 91.30% in study group was significantly higher than the control group 69.57%, the difference was statistically significant(χ2 =5.874, P=0.015).The serum levels of TNF-α, IL-2 and IL-6 in study group were significantly lower than the control group, the difference was statistically significant(P<0.05).The levels of CD3 +, CD4 +, CD8 +and NKT in study group were significantly higher than the control group, the difference was statistically significant(P<0.05).Followed up for 1 year, the recurrence rate 17.78%(8/45) in study group was significantly lower than the control group 46.67%(21/45), the difference was statistically significant(χ2 =8.598, P =0.003).Conclusion The Shenling Baizhu power treatment of recurrent aphthous ulcer has better clinical curative effect, can reduce levels of inflammatory factor, improve immune function, and reduce the recurrence rate.
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Objective To analyse the effects of high insulin on the expression and function of P-glycoprotein (P-gp), and preliminarily investigate the influence of insulin on chemotherapeutic sensitivity in MCF-7/ADR cells. Methods MCF-7/ADR cells were cultured with different concentrations of insulin(0.001, 0.005, 0.01, 0.05 and 0.1μmol/L). Real-time PCR was used to detect the expression of P-gp mRNA. Western blot assay was used to detect the expression level of P-gp. Rhodamine 123 was used to detect the efflux function level of P-gp. Cell viability and chemotherapeutic sensitivity were detected by MTT assay. Results High concentration of insulin (0.1 μmol/L) promoted the proliferation of MCF-7/ADR cells. The concentration of insulin (0.05 and 0.1 μmol/L) accelerated P-gp mRNA and protein expression, which also augmented the efflux function of P-glycoprotein and reduced the chemotherapeutic sensitivity to epirubicin. Conclusion High concentration of insulin may influence the drug resistance of breast cancer cells by promoting the expression and function of P-glycoprotein of MCF-7/ADR cells.
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Objective To investigate the clinical features, phenotypes and genotypes of Pseudomonas aeruginosa (PA) strains isolated from patients with diabetic foot infection (DFI) resisting to aminoglycosides antibiotics (AmAn). Methods The clinical profiles of 209 DFI patients hospitalized in the Tianjin Metabolic Diseases Hospital were collected and ana?lyzed. Forty-one PA strains were identified, and their antibiotic resistance profiles were obtained. The DNAs of PA isolates were extracted and applied to amplifications for several aminoglycosides modifying enzyme genes, including aac(3′)-Ⅰ, aac (3′)-Ⅱ, aac(6′)-Ⅰb, aac(6′)-Ⅱ, ant(2′′)-Ⅰand ant(3′′)-Ⅰby PCR method. Combining with the clinical features and the antibiotic resistance profiles, the relationship between genotypes and phenotypes of the PA strains was analyzed. Results Gram positive bacteria (G+) were the majority of the pathogen with 51.67%detection rate. The total detection rate of PA was 19.62%, listed as the top one pathogenic bacterium among gram negative bacteria (47.67%). There was significant difference in the ratio of ulcer area≥4 cm2 between PA group and non-PA group and G+group. There were significantly higher inci?dence rate of ischemic ulcer and osteomyelitis in PA group than those of G+group. There were higher clinical characteristics and ulcer depth (SAD) score, and increased hypersensitive C-reactive protein in PA group than those of G+ group. There were 30 strains of PA being resistant to AmAn (73.17%). The predominant drug resistance gene to AmAn was ant(3′′)-Ⅰ(65.85%), and aac(3′)-Ⅰgene was not found from all PA isolates. Conclusion The detection rate of PA isolated from DFI patients was higher, and patients were with the characteristics of larger, deeper and severe ischemia of ulcer area. The phe?nomenon of PA resistant to AmAn was more serious, and ant(3′′)-Ⅰgene identified from PA isolates was the most common resistance gene identified to AmAn.
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Objective To observe the effects of high glucose and anoxia on Amot expression in vascular endothelial cells (VECs),and explore its role in angiogenesis.Methods VECs were incubated with different glucose concentrations for 48 h,and then cultured at normal oxygen concentration or anaerobic condition for 24 h.The protein expressions of p130-Amot and p80-Amot were detected by Western blot.After Amot expression was downregulated in VECs by siRNA,wound healing experiments and angiogenesis experiments were performed to test the effect of decreased Amot expression on angiogenesis.Results pl30-Amot protein expressions in low glucose (5.5mmol/L) plus normal oxygen group and low glucose plus anaerobic group were higher than those in high glucose (30mmol/L) plus normal oxygen group,high glucose plus anaerobic group,middle glucose (15 mmol/L) plus normal oxygen group,and middle glucose plus anaerobic group (all P＜0.01).Compared with low glucose plus anaerobic group,p130-Amot expression was higher in low glucose plus normal oxygen group (P ＜ 0.01).However,the expression of p80-Amot showed no statistically significant difference among different groups (P＞0.05).Compared to the normal VECs,the cells with decreased Amot expression by siRNA exhibited an attenuated cell migration in the wound healing experiments and a lesser tube formation in the angiogenesis experiments.Conclusions High glucose exerts a more significantly negtive effect on the Amot expression than anoxia in VECs.The downregulation of Amot expression inhibits migration and angiogenesis of VECs.
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<p><b>BACKGROUND</b>Copious evidence from epidemiological and laboratory studies has revealed that sleep status is associated with glucose intolerance, insulin resistance, thus increasing the risk of developing type 2 diabetes. The aim of this study was to reveal the interaction of sleep quality and sleep quantity on glycemic control in patients with type 2 diabetes mellitus.</p><p><b>METHODS</b>From May 2013 to May 2014, a total of 551 type 2 diabetes patients in Tianjin Metabolic Diseases Hospital were enrolled. Blood samples were taken to measure glycosylated hemoglobin (HbA1c), and all the patients completed the Chinese version of the Pittsburgh Sleep Quality Index (PSQI) questionnaire to evaluate their sleep status. "Good sleep quality" was defined as PQSI <5, "average sleep quality" was defined as PQSI 6-8, and "poor sleep quality" was defined as PQSI >8. Poor glycemic control was defined as HbA1c ≥7%. Sleep quantity was categorized as <6, 6-8, and >8 hours/night. Short sleep time was defined as sleep duration <6 hours/night.</p><p><b>RESULTS</b>In the poor glycemic control group, the rate of patients who had insufficient sleep was much higher than that in the other group (χ(2) = 11.16, P = 0.037). The rate of poor sleep quality in poor glycemic control group was much greater than that in the average control group (χ(2) = 9.79, P = 0.007). After adjusted by gender, age, body mass index, and disease duration, the adjusted PSQI score's OR was 1.048 (95% CI 1.007-1.092, P = 0.023) for HbA1c level. The sleep duration's OR was 0.464 (95% CI 0.236-0.912, P = 0.026) for HbA1c level. One-way analysis of variance showed that the poor sleep quality group had the highest homeostasis model assessment-insulin resistance (P < 0.01).</p><p><b>CONCLUSIONS</b>Inadequate sleep, in both quality and quantity, should be regarded as a plausible risk factor for glycemic control in type 2 diabetes. Poor sleep might bring much more serious insulin resistance and could be the reason for bad glycemic control. A good night's sleep should be seen as a critical health component tool in the prevention and treatment of type 2 diabetes. It is important for clinicians to target the root causes of short sleep duration and/or poor sleep quality.</p>
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Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Blood Glucose , Metabolism , Physiology , Cross-Sectional Studies , Diabetes Mellitus, Type 2 , Blood , Sleep , PhysiologyABSTRACT
Objective To investigate the clinical features and antibiotic susceptibility of osteomyelitis infected by Gram-negative bacteria (G-) in patients suffered from diabetic foot ulcers (DFU). Methods The clinical data of 91 DFU pa-tients accompanied with osteomyelitis (DFO) were retrospective studied. These patients hospitalized in the Tianjin Metabolic Diseases Hospital were divided into two groups, Gram-negative bacteria (G-) group (n=44) and Gram-positive bacteria (G+) group (n=42), respectively. The clinical features were compared between two groups. Logistic regression analysis was used to determine the risk factors for Gram-negative bactreial infection. The Gram-negative antibiogram was summarized. Results A total of 112 pathogens were isolated from 91 patients. G-bacteria were the most frequent pathogens (48.2%), following by G+ bacteria (47.3%) and fungi (4.5%). Pseudomonas aeruginosa was the majority of the G-bacteria. Comparing the two groups, the rate of antibiotic use within the previous 6 months was significantly higher in G-group (75.0%) than that of G+group (52.4%, P<0.05). There were no significant differences in the other indicators between two groups. The Logistic re-gression analysis revealed that the history of antibiotic use was the independent risk factor of G-bacterial infections in DFO patients. Antibiotics susceptibilities reflected G- bacteria were more prevalent to resist to cephalosporins and quinolonem, but sensitive to imipenem, ceftazidine and cefperazone-sulbactam. Conclusion Gram negative bacteria were not only the main pathogens isolated from DFO patients, but also frequently resistant to several popular antibiotics in China. The proper bacteria culture and antibiotic sensitivity test are especially emphasized to patients with DFU.
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Objective To investigate clinical features and antibiotic resistance of pseudomonas aeruginosa (PA) strains isolated from patients with diabetic foot infections (DFI) in Tianjin Metabolic Diseases Hospital.Methods Eighty-five PA strains were isolated from 428 patients with diabetic foot in the hospital from Jan 2008 to Dec 2010.The clinical features of patients were summarized.Relationships between the isolates and depth of ulcer or severity of infection were analyzed.The disk-diffusion method was performed to examine antimicrobial susceptibility.Results Gram positive (G+) and Gram negative (Gˉ) isolates were 50.47％ and 41.12％,respectively.Multidrug-resistant PA composed 32.9％ of the total PA isolates.The size of ulcers with PA infections was bigger than those with non-PA bacterial infections (P＜0.05).Compared to G+ strains,patients with PA strains were older,had lower hemoglobin,but higher serum sensitive C-reactive protein; and more frequently,they had ischemic ulcer and osteomyelitis.Compared to G+ strains,the PA strains were more frequently isolated from deeper ulcers and with more serious infections(P＜0.05).The resistant rates of PA to cephalosporins,fluoroquinolones,and aminoglycosides were between 32.9％-61.2％,37.6％-42.4％,and 37.6％-62.4％,respectively.Only one out of 85 PA strains was imipenem-resistant.However,sensitiveness of all PA isolates to cefoperazone and sulbactam reached 100％.Conclusion PA strains are mainly found in patients with deeper ulcers and more serious infections.Multidrug-resistant PA is common in DFI.It is important to isolate pathogens and determine their antibiotic resistance correctly in diabetic foot patients in order to provide appropriate drug administration and to reduce the production and dissemination of drug resistant strains.
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Objective To investigate the infection and risk factors of multidrug-resistant Pseudomonas aeruginosa in diabetic foot.Methods Totally 85 strains of Pseudomonas aeruginosa were isolated from 428 samples of diabetic foot ulcers in Tianjin Metabolic Diseases Hospital from Jan 2008 to Dec 2010.Drug sensitivity tests were performed and multivariate logistic regression analysis was used to examine the risk factors of multidrug-resistant Pseudomonas aeruginosa infection. Results In 85 strains of Pseudomonas aeruginosa, 28 (32.94％ ) were multidrug resistant. Multidrug-resistant Pseudomonas aeruginosa showed high resistance to 3rd generation cephalosporins,quinolones and aminoglycosides with resistant rates of 42.86％-67.86％,32.14％ -57.57％,and 42.86％-67.86％,respectively. Logistic analysis indicated that previous antibiotic treatments, hypertension and anemia were associated with multidrug-resistant Pseudomonas aeruginosa infection ( OR =5.758,0.257 and 0.270,P =0.006,0.014 and 0.013 ). Conclusion Multidrug-resistant Pseudomonas aeruginosas isolated from diabetic foot are highly resistant to several commonly used antibiotics,and previous antibiotics use,hypertension and anemia are risk factors for multidrug-resistant Pseudomonas aeruginosa infections.
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Objective To investigate SCCmec genotypes and drug-resistance profiles of the methieillin-resistant Staphylococcus epidermidis (MRSE) strains isolated from the patients suffered from diabetic foot infections (DFI) in the Tianjin Metabohc Diseases Hospital. Methods After dabridement, specimens of 390 infectious diabetic foot ulcers in the hospital from Jan 2008 to Jun 2010 were collected from the wound basal parts by cotton swab for culture. The disk-diffusion method was performed to examine antimicrobial susceptibility. DNAs of the MRSE strains were extracted, and their SCCmec genotypes were identified by PCR. Results Twenty of the seventy(28.6% ,20/70)Staphylococcus epidermidis strains were mecA posifive. Among the MRSE isolates, 2 ( 10.0% )were SCCmec Ⅱ ,9 (45.0%)were SCCmecⅢ and 9 (45.0%)were SCCmec Ⅳ. None of the isolates were genotyped as SCCmec Ⅰ or Ⅴ. No mater which genotypes they were, all the MRSE isolates were multi-drug resistant. They were resistant not only to β-lactams (including penicillins, cefoxitin and cephems), but also to non-β-lactams (including macrolides, fiuoroquinolones and sulfonamides ) . Resistance to voncomycin and rifampicin were not found in these strains . Conclusion SCCmec Ⅲ and SCCmecⅣ are major genotypes of the MRSE isolates from the infectious diabetic foot ulcers.The SCCmec Ⅳ genotype strains with multi-drug resistant profiles are prevalent in the diabetic foot infections.
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Obiective:To investigate the effect of interleukin-6(IL-6)on apolipoprotein(apo)M expression in a human hepatoblastoma cell line(HepG2).Methods:HepG2 cells were cultured and incubated with different concentrations of IL-6 (0,1.25,2.5,5,10,20,40 and 80 μg/L)for 24 hours.After the incubations,total RNAs were extracted and applied to reverse transcript PCR and real-time quantitative PCR to detect the expression levels of apoM and apoA-I.The effect of IL-1α on apoM expression was also determined.Results:IL-6 significantly inhibited the expression of apoM(F=10.778,P ＜ 0.01).Whereas IL-6 did not influence the expression of apoA-I(F=2.004,P ＞ 0.05).IL-1α(0,1.25,2.5,5,10,20,40 and 80 μg/L)did not decrease the expression of apoM(F=2.038,P ＞ 0.05).Cooclusion:IL-6 inhibits apoM mRNA transcription,which may contribute to the pathogenesis of abnormal lipid metabolism and macrovascular complications in type 2 diabetes.
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Objective To prepare recombinant human epidermal growth factor (rhEGF) sustained-release microspheres and evaluate their morphology, rhEGF releasing activities and cell proliferation activity in vitro and compare difference of rhEGF sustained-release microspheres and rhEGF in facilitaring ulcer healing in diabetic rats. Methods (1) rhEGF sustained-release microspheres were prepared by the modified double emulsion method. Morphology of the microspheres was detected by transmission electron microscope and size distribution measured by laser granularity meter/Zeta electric potential meter. ELISA assays were applied to determine rhEGF releasing. (2)Proliferation of mouse fibroblasts was analyzed by MTr method. (3) Diabetic rat models were prepared and divided into four groups, ie, rhEGF sustained-release mierospheres group (Group A), rhEGF stock solution group (Group B), blank sustainedrelease mierospheres group (Group C) and PBS meustruum control group (Group D), which were given drug once a day. The wound healing rate was calculated by taking photographs at days 3,7,14 and 21. Skin specimens from the wound edge were harvested partially for observation of hydroxyproline (HYP) contents. Immunohistochemistry was employed to detect integrin 131 and keratin-19 and measure their positive staining area ratio. Results (1) The particle diameter of rhEGF sustained-release microspheres was 193.5 nm, with relative uniform particle diameter distribution. There showed no conglutination among rhEGF susrained-release microspheres, with good dispersibility. Releasing drug lasted for 24 hours and accorded with Higuchi release kinetic model. (2) Different concentrations of rhEGF sustained-release microspheres could promote the proliferation of mouse fibroblast, especially the concentration of 10 μg/L (P <0.05, compared with the control). (3) From the 7th day after treatment, Group A had the fastest wound healing rate, with statistical difference compared with other three groups (P < 0.05). Group A had higher HYP contents and positive area ratio of integrin β1 and keratin-19 than Group B. Conclusions rhEGF sustained-release microspheres prepared by the modified double emulsion method have uniform particle size and can last release for 24 hours. Compared with rhEGF stock solution, rhEGF sustained-release microspheres have faster and better ulcer healing and higher healing quality in diabetic rats.