Objective To explore the potential impact of low concentration of metformin on the morphology and function of mitochondria of HepG2 cells. Methods HepG2 cells in experimental group and control group were treated with or without low concentration of metformin (1mM/L), respectively. The cells were incubated for 12h in the incubator with constant temperature and humidity as well as 1% oxygen. Orange Mitoview was used to stain the mitochondria to detect the effects of the drug on its morphology and quantity. Transmission electron microscope was utilized to observe the effect of metformin on the ultrastructure of mitochondria. The mitochondrial respiratory chain complex I activity in HepG2 cell was detected by Complex I Enzyme Activity Dipstick Assay Kit (DAK). Results Orange Mitoview staining showed that low concentration of metformin had little effect on the morphology and number of mitochondria of cells in experimental group , and the difference between control and experimental group was not statistically significant (P > 0.05). In addition, the result was further determined by transmission electron microscopy. However, DAK analysis showed that complex I activity of cells in experimental group was significantly lower than that in control group. Conclusion Under Hypoxia conditions, low concentration of metformin had no significant effect on the morphology and number of mitochondria of HepG2 cells, but it significantly reduces the activity of mitochondria of HepG2 cells.

Objective To investigate the mechanism and epidemiological characteristics of carbap-enem-resistant Proteus mirabilis ( PM) strains deficient in swarming motility. Methods PM strains were isolated from Hangzhou General Hospital of CAPF ( Chinese People′s Armed Police Forces) during January 2013 to December 2014. Bacterial motility and flagella of the PM strains were observed through semi-solid agar culture and flagella staining. Pulsed-field gel electrophoresis ( PFGE) was performed for homology anal-ysis. Antimicrobial susceptibility test and phenotypic confirmatory test were also carried out. PCR analysis and DNA sequencing were performed to confirm the genotype of resistant genes. Plasmid electroporation and S1-PFGE in combination with Southern blot hybridization were used to determine the location of the carbap-enem-resistant genes. Genetic structure of the blaKPC-2 gene was obtained by PCR mapping. Results A total of 42 PM isolates deficient in swarming motility were screened out and the resistance rates to imipenem and meropenem were 57. 1% and 52. 4%, respectively. PCR analysis and DNA sequencing confirmed that 24 carbapenem-resistant PM isolates deficient in swarming motility carried blaKPC-2 gene and belonged to three clones as indicated by the results of PFGE. Southern blot hybridization indicated that the blaKPC-2 gene was located on plasmids varying in size (26 kb, 55 kb and 139 kb). In addition, some of the strains harbored several resistant genes, such as blaTEM-1 , blaCTX-M-65 and rmtB. The genetic structures of strains carrying blaKPC-2 gene were ISKpn8, blaKPC-2 and ISKpn6-like from upstream to downstream. Conclusion Compared with the PM strains with swarming motility, the carbapenem-resistance rate was significantly higher in these PM strains deficient in swarming motility. Carbapenemases KPC-2 played an important role in the carbapen-em-resistant PM strains deficient in swarming motility. There was a cloning spread trend for carbapenem-re-sistant PM strains in our hospital. Clinicians should pay more attention to the risk of spreading.

Objective To evaluate the usefulness of serum galactemannan(GM) for the diagnosis of invasive pulmonary aspergillosis (IPA) in critically ill patients. Methods Study was conducted between February 2007 and July 2008. Included patients on admission ICU who suffer from suspected IPA. GM test and culture were collected 2 weekly. Chnical feature, mycological evidence and optical density index (ODI) were noted. Clinically invasive fungal infection(IFI) were defined proven, probable and possible. The patients were classified into neutropenia, non-neutropenia and treated with immunosuppressive agents, non-neutropenia and non-immunosuppressive agents. To compared of the sensitivity and specificity of GM in different patients. Results 94 patients were included, 4 patients were proven, 29 patients were probable, 34 patients were possible IFI, 27 patients were non-IPA. The positive rate of the GM was 31.9% (30/94). The sensitivity and specificity of GM in proven cases and probable cases are 66.7% and 92.6%. GM assay tended to become positive earlier than the culture 2-10(5.33±2.17)d. We found that differences in patient diagnosis and selection might account for the disparities seen for positive rate for the GM test. There was positive in three of the four patients with proven, the positive rate of GM was 65.5% for probable cases, for possible cases was 17.6%, for non-IPA cases was 7.4% (P=0.001). For patient with neutropenia , treated with immunosuppressive agents and without immunosuppressive agents, the positive rate of GM was 52.9%vs 41.7% vs 34. 6% (P=0.015) ;the sensitivity was 80.0% vs 70. 0% vs 53.8% (P=0.011), the ODI was 1.365 (0.582-6.736) vs 1. 123 (0. 623-6.868) vs 0.554 (0.522-0.823), P=0. 005, respectively. Conclusion These results show that GM test is useful for early diagnosis IPA in critically ill patients. Differences in patient selection and diagnosis might account for the disparities seen for positive rate and sensitivity for the GM test. It has been higher sensitivity and ODI in the patient treated by immunosuppressive agents.

Objective To investigate the clinical role of central venous pressure(CVP) to evaluate fluid responsiveness in septic shock patients. Methods 66 septic shock patients were studied, every patient was administered a volume challenge, before and after it, CVP, intrathoracic blood volume index (ITBVI),global end-diastolic volume index(GEDVI), cardiac index(CI), stroke volume index(SVI) were measured by PiCCO method. All the obtained values were analyzed by statistics method. Results Initial CVP in responders is significantly different from that in nonresponders; △ITBVI, △GEDVI, △CI, △SVI, △HR (△:changes) before and after volume challenge in responders were significantly different from those in nonresponders; the significance of △ITBVI, AGEDVI to predict volume responsiveness was strong indicated by high values of areas under the receiver operating characteristic curves (0.674 and 0.700, respectively).If patients were regrouped by CVP≤11 mm Hg(1 mm Hg=0.133 kPa) and CVP > 11 mm Hg, initial ITBVI and GEDVI in responders were not significantly different from that in nonresponders; △ITBVI,△GEDVI, △CI, △SVI before and after volume challenge in responders were significantly different from those in nom'esponders. Conclusion In septic shock patients, CVP play a guidance role to predict and evaluate volume responsiveness and when CVP was > 11 nun Hg, a positive response will be less likely. Initial volumetric parameters(intrathoracic blood volume and global end-diastolic volume) play a questionable role in predicting and evaluating volume responsiveness, changes before and after volume challenge maybe helpful.

Objectlve To research and analyze the hemod)rnamic status of refractory septic shock associated cardiac dysfunction.Methods 70 refractory septic shock patients were studied.In the duration of pulmonary artery catheter(PAC)-directed hemodynamic optimization,the patients were divided into a cardiac dysfunction group and a control group.Hemodynamic parameters,arterial blood lactate concentration and APACHE II scores were obtained instantly after the placement of a PAC,then lactate clearance in 24 hours was surveyed and calculated.Subsequently the two groups of patients were regrouped by nonsurvivor and survivors respectively.All the obtained values were analyzed with statistic methods.Results 37% of the refractory septic shock patients was complicated with cardiac dysfunction.The age of the patients complicated with cardiac dysfunction was significantly higher than that of the patients of the control group.Central venous pressure(CVP),pulmonary artery obstruction pressure(PAOP),pulmonary artery pressure (PAP),systemic vascular resistance index(SVRI),pulmonary vascular resistance index(PVRI)and oxygen extraction ratio(O2ext)in the cardiac dysfunction group were significantly different from those in the control group.Cardiac output(CO),cardiac index(CI),oxygen delivery index(DO2I)and mixed venous oxygensaturation(S-v O2)were significantly lower than those of the patients in the control group.S -v O2 had a strong correlation witIl CI.If the patients were regrouped by nonsurvivors and survivors.in the patients complicated with cardiac dysfunction APACHE II scores were significantly higher in the nonsurvivors than survivors:the lactate clearance in 24 hours(median-25%)of the nonsurvivors was significantly lower than that of nonresponders(median 22%),P＜0.05.Conclusion (1)In refractory septic shock patients,cardiac dysfunction maybe the main reason leading to bad outcome.(2)Higher CVP and PAOP and lower S -v O2 indicate the onset of cardiac dysfunction.(3)The patients with significantly high initial arterial blood lactate level and the low lactate clearance in 24 hours had bad outcome.

Objective To evaluate the effects of curcumin on the expression of receptor for advanced glycation end-products (RAGE) in the spinal dorsal horn and dorsal root ganglion (DRG) of the rats with type 2 diabetic neuropathic pain (DNP).Methods Male Sprague-Dawley rats,weighing 160-180 g,were used in this study.Type 2 diabetes mellitus was induced by high-fat and high-sucrose diet for 8 weeks and intraperitoneal streptozotocin (STZ) 35 mg/kg and confirmed by fasting blood glucose level ≥ 16.7 mmol/L 3 days later.Type 2 DNPwas confirmed by the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) measured on day 14 after STZ administration less than 80％ of the baseline value.Eighty-one rats with type 2 DNP were randomly divided into 3 groups (n =27 each) using a random number table:DNP group,DNP + curcumin group (DCur group),and DNP+ solvent group (group DSC).In DCur and DSC groups,curcumin 100 mg· kg-1 · d-1 and corn oil 4 ml · kg-1 · d-1 were injected intraperitonally,respectively,for 14 consecutive days starting from the day 14 after STZ administration.Another 27 normal male Sprague-Dawley rats served as control group (group C) and were fed with normal forage.MWT and TWL were measured before STZ injection,at day 14 after STZ injection,and on 3,7 and 14 days after curcumin injection.RAGE positive cells were determined by immuno-histochemistry and the expression of RAGE by Western blot in the spinal dorsal horn and DRG after MWT and TWL were measured on 3,7 and 14 days after curcumin injection.Results Compared with group C,MWT was significantly decreased and TWL was shortened at 14 days after STZ injection and each time point after curcumin injection,the rate of RAGE positive cells in the spinal dorsal horn and DRG was increased at each time point after curcumin injection,and the expression of RAGE was up-regulated in the spinal dorsal horn at each time point after curcumin injection and in the DRG at 7 and 14 days after curcumin injection in group DNP.Compared with group DNP,MWT was significantly increased and TWL was prolonged at each time point after curcumin injection,the rate of RAGE positive cells in the spinal dorsal horn and DRG was significantly decreased at 7 and 14 days after curcumin injection,and the expression of RAGE was down-regulated in the spinal dorsal horn at each time point after curcumin injection and in the DRG at 7 and 14 days after curcumin injection in group DCur,and no significant change was found in the parameters mentioned above in group DSC.Conclusion The mechanism by which curcumin attenuates DNP may be related to inhibition of up-regulated expression of RAGE in the spinal dorsal horn and DRG of rats with type 2 diabetes mellitus.

Objective To evaluate the effect of curcumin on the expression of MMP-2 and MMP-9 in the spinal dorsal horn and dorsal root ganglion (DRG) of rats with type 2 diabetic neuropathic pain (DNP).Methods Type 2 diabetes mellitus was induced by high-fat and high-sucrose diet and intraperitoneal streptozotocin 35 mg/kg,and confirmed by fasting blood glucose level ≥ 16.7 mmol/L in male Sprague-Dawley rats.Type 2 DNP was confirmed by the mechanical paw withdrawal threshold (MWT) and thermal paw withdraw latency (TWL) measured on day 14 after streptozotocin administration ＜ 80％ of the baseline value.The rats were then randomly divided into 3 groups (n =27 each):type 2 DNP group (group DNP); curcumin group (group Cur); solvent control group (group SC).In Cur and SC groups,curcumin 100 mg/kg and corn oil 4 ml/kg were injected intraperitonally,respectively,once a day for 14 consecutive days starting from day 14 after streptozotocin administration.Another 27 normal Sprague-Dawley male rats served as control group (group C) and were fed with normal forage.MWT and TWL were measured before type 2 DNP was induced,after type 2 DNP was induced,and at 3,7 and 14 days after curcumin injection(T1-5).The rats were sacrificed after MWT and TWL were measured at T3-5,and the lumbar segments of the spinal cord and DRG (L4-6) were removed for determination of the expression of MMP-2 and MMP-9 by Western blot.Results Compared with group C,MWT was significantly decreased and TWL was shortened,and MMP-2 and MMP-9 expression in the spinal dorsal horn and DRG was up-regulated in DNP,Cur and SC groups.Compared with DNP group,MWT was significantly increased and TWL was prolonged,and MMP-2 and MMP-9 expression in the spinal dorsal horn and DRG was down-regulated in Cur group,and no significant changes were found in the parameters mentioned above in SC group.Conclusion The mechanism by which curcumin attenuates type 2 DNP may be related to up-regulation of the expression of MMP-2 and MMP-9 in the spinal dorsal horns and DRG of rats.

Objective To investigate the effects of hemolysis on glycated albumin (GA) determined by ketoamine oxidase method and its correction.Methods GA concentration and hemolytic parameter(optical density,A) in non-hemolytic serum and different degree hemolytic serum samples were measured.The impact of hemolysis on GA and the relationship between hemolysis and GA were analysed.A formula was developed to correct the interference of hemolysis on GA measurement using regressive Multiple analysis.Results Compared with non-hemolytic serum,hemolysis resulted in the significantly decreased concentrations of GA detected by ketoamine oxidase method(P ＜ 0.01),which were significantly associated with the degree of hemolysis (R2 =0.943 4).Y and Z represented GA concentration of non-hemolytic serum and different degree hemolytic serum,while X represented optical density of hemolytic parameter.Formulas for GA measurement were presented:Y =2.468X + Z-0.015 73,GA concentrations of hemolytic samples can be reverted to the values without statistical difference from the GA concentration in corresponding non-hemolytic samples.The bias of corrected GA was less than 10％.Conclusion Our results indicate that the level of GA measured through ketoamine oxidase method is negatively affected by hemolysis.The formula of mathematical correction of GA results in hemolytic samples should be suitable for the requirements of clinical laboratory.

Objective:To investigate the correlation between IL-10 and I1-17 expression levels in Mycoplasma pneumoniae pneumonia (MPP) and lung function.Methods:70 patients were included in this study.According to wheezing or not,they were divided into wheezing group and non-wheezing group.Another 30 healthy children were taken as a control group.After taking fasting blood 5ml,the serum IL-10 and IL-17 expression levels were detected by ELISA.The forced expiratory volume in one second (PEV1),peak expiratory flow (PEF) and forced vital capacity (FEV1 / FVC) of all subjects were detected.Results:The IL-10 expression level of the wheezing group were significantly different with that of the control group (P＜0.05) and that of the non-wheezing group (P＜0.05).The 1L-17 expression level of the wheezing group also had significant difference (P＜0.05) with that of the control group and non-wheezing group (P＜0.05).The IL-10 expression levels of wheezing and non-wheezing group all were lower than that of the control group.Whereas the IL-17 expression levels of wheezing and non-wheezing group all were higher than that in the control group.In addition,patients in wheezing group had higher PEV1,PEF,PVE1/FVC values than those in non-wheezing group,with significant difference (P＜0.05).The serum level of IL-10 expression of Mycoplasma pneumonia patients was positively correlated with PEV1,PEF and PVE1/FVC,while the serum level of IL-17 expression of Mycoplasma pneumonia patients was negatively correlated with PEV1,PEF and PVE1/FVC.Conclusion:The serum levels of IL-17 and IL-10 expression of Mycoplasma pneumonia children had close correlation with their pulmonary function.

Objective The aim of this study was to evaluate whether PRDM16 gene polymorphisms were associated with dyslipidemia. Methods The polymorphisms of rs2651899, rs2236518, rs870171, and rs2282198 in PRDM16 gene in 528 participants were genotyped by the method of snapshot or ligase detection reaction. The genotype differences and the allele differences between the case group and the control group were analyzed. Linkage disequilibrium analysis was performed with SHE-sis online software. The interaction between rs2651899, rs2236518, rs870171, rs2282198 and gender, age, BMI were analyzed by MDR software. Results The frequency of allele A in rs2651899 locus was significantly higher in low HDL-C group compared with that in control group[OR(95%CI)=1.32(1.02-1.71), P=0.033]. The frequency of A/C genotype in rs870171 was significantly different between LDL-C abnormal group and control group[OR(95% CI)=1.97(1.01-3.86), P=0.037]. There may be interaction between rs2236518 and sex, which is a risk factor for low HDL-C[Model Ⅱ: OR(95% CI)=1.958(1.366-2.809), P<0.01]. There may be interactions among rs2651899, rs2236518, rs870171, and rs2282198, which seemed to be risk factors for lower HDL-C[Model Ⅳ: OR(95% CI)=3.991(2.707-5.884), P<0.01]. rs870171, rs2282198 may have interaction with age, which is a risk factor for high LDL-C [Model Ⅶ: OR(95%CI)=3.991(2.707-5.884), P<0.01]. Conclusion Allele A of rs2651899 may be a risk factor to low HDL-C. Under the codominant inheritance patterns, genotype A/C of rs870171 may be a risk factor to high LDL-C. In addition, there may be interaction between SNPs with gender and age.