Objective To investigate the effect of RBP on 3T3-L1 adipocyte differentiation,lipid metabolism and glucose transporter 4(GLUT-4)gene expression.Methods We constructed an expression vector for rat resistin gene and transfected it into 3T3-L1 adipocytes.RBP was added to the medium of 3T3-L1 adipocytes or resistin-overexpressing adipocytes on day 0 of differentiation.Cell differentiation and lipid accumulation were determined by oil red O staining.The mRNA expressions of differentiation marker genes(pref-1,C/EBP?,FAS)and GLUT-4 gene were evaluated by RT-PCR.Triglyceride(TG)and free fatty acids(FFAs)in adipocytes were measured by colorimetric kit.Results(1)When 10-12mol/L RBP was applied,the percent of living cells was high and the shape was unchanged.(2)RBP had no effect on the differentiation of normal adipocytes,but significantly decreased the number of lipid droplets in resistin-overexpressing adipocytes without affecting the lipid droplets-presenting day.(3)C/EBP? and FAS expressions in resistin-overexpressing adipocytes were down-regulated after RBP was applied,without changing their expressions in normal adipocytes.(4)RBP had no effect on the cellular TG and FFAs levels in normal cells,whereas it can significantly decrease the levels in resistin-overexpressing adipocytes.(5)There was no difference in the expression of GLUT-4 gene between 3T3-L1 adipocytes and RBP-applied cells.Conclusions(1)RBP has no effect on the cell differentiation and lipid metabolism in normal 3T3-L1 adipocytes.(2)RBP can inhibit the cell differentiation and lipid metabolism of resistin-overexpressing 3T3-L1 cells.(3)RBP has no effect on the expression of GLUT-4 gene.

Objective To analyze the distribution and antimicrobial resistance of pathogenic bacteria in urinary tract infection (UTI)so as to provide evidence for appropriate selection of antimicrobial agents in clinical practice. Methods From January 2001 to December 2008 in Shanghai Ruijin Hospital,4683 strains of pathogenic bacteria isolated from urine samples were detected by ATB system;drug susceptibility test was performed with disk diffusion method and pathogenic bacteria distribution and drug resistance was analyzed with WHO NET 5.3 software. Results Among 4683 strains of pathogenic bacteria,most was gramnegative bacilli,accounting for about 77.8%,of which predominant strain was Escherichia coli (68.7%,3217/4683).The predominant strain of gram-positive bacteria was Enterococcus faecalis,accounting for 10.0%(468/4683).Escherichia coli showed hish resistance rotes to ampicillin,piperacillin and compound snlfamethoxazole(SMZ-TMP),which were 76.6%,61.7%and 57.4%respectively,while a low resistance to imipenem,cefoperazone-sulbactam,piperacillin-tazobactam,Enterococcus faecalis showed high resistance rates to erythromycin,gentamicin and levofloxacin,which were 65.8%,43.2%and 31.1%respectively,and were most susceptive to vancomycin and teicoplanin, both with resistance rates of 0. The susceptibility rate of Enterobacteriaceae to imipenem was 100%. From 2006 to 2008, the detection rate of extend-spectrum β-lactamases ESBLs -producing Escherichia coil in outpatient increased year by year, from 28.7% to 43.3% (P＜0.05), whereas no significant change was found in inpatients. The detection rate of (ESBLs)-producing Escherichia coil in inpatients was significantly higher than that in outpatients (P＜0.05).The detection rate of ESBLs-producing Escherichia coil was 23.6%. The antimicrobial resistance rate in elderly patients was significantly higher than that in overall antimicrobial resistance rote (P＜0.05). Conclusions The predominant bacteria of UTI are still gram-negative bacteria, main of which is Escherichia col. Bacteria are resistant to a variety of antibiotics. Approximate selection of antibiotics in clinical practice should be made on the basis of susceptibility test results.

Objective To investigate the application value of montelukast in patients with stable chronic obstructive pulmonary disease (COPD).Methods 64 patients with stable COPD in Jinqiu Hospital of Liaoning Province from December 2015 to December 2016 were randomly divided into observation group and control group according to the digital table,32 cases in each group.The two groups were given routine treatment,including cough,phlegm,oxygen therapy,smoking cessation,rehabilitation treatment,regular use of long-acting bronchodilators,on this basis,the observation group was treated with montelukast sodium.After 12 weeks of treatment,the patients were assessed for airway improvement and quality of life before and after treatment,and recorded the two groups of acute exacerbation and adverse drug reactions.Results The two groups had improved lung function and quality of life after treatment,but after treatment,the FEV1,FVC and FEV1 actual/FEV1 prediction of the observation group were significantly higher than those of the control group,the differences were statistically significant (t =2.49,1.77,1.85 all P ＜ 0.05).After treatment,the 6 minutes walking distance of the observation group was longer than that of the control group,Borg score of the observation group was lower than that of the control group,the differences were statistically significant (t =1.83,6.66,all P ＜0.05).After treatment,the SGRQ score in the observation group was lower than that of the control group,the difference was statistically significant[(51.15 ± 3.14)points vs.(54.83 ± 4.03)points,t =3.94,P ＜ 0.01].Conclusion Montelukast can effectively improve the stability of COPD pulmonary function and quality of life,and with high safety.

Objective To investigated the inhibitory effect of an IgG-Fc region specific inhibitory peptide on the ANCA-accelerated apoptosis of neutrophils. Methods The peptide was prepared by solid-phase peptide synthesis and its biological activity was identified by rosette formation assay. ANCA was prepared from the sera of active Wegener's granulomatosis (WG) and microscopic polyangiitis (MPA) patients. Neutrophils isolated from the blood of healthy volunteers were primed with TNF-?(2 ng/ml) then incubated with ANCA. At different intervals(3, 6, 12, 18 hours) the neutrophils were harvested to assess the apoptosis by flow cytometric analysis of JC-1 staining, Sub-G1 population and fonnation TUNEL technique. Results Tg19320 bound tightly to human IgG dose-dependendy and inhibited statistically the rosette formation between SRBC-IgG and U937 cells(20.3% vs 53.2% ,P

The aim of the study was to observe osseointegration of dental implants in osteoporotic rats. Rats were randomly divided into groups A(SHAM) and B(OVX), and pure titanium screws were implanted in the distal half of the right femurs. 1 5 and 3 months after implantation, half of the rats in each group were sacrificed and right femurs were observed by X ray photography, scan electron microscopy as well as light microscopy. The results showed that 1.5 and 3 months after implantation, the osseointegration index (OI) of group A was higher than that of group B. But the OI of each group was almost the same at different intervals. 1 5 months after implantation, the amount of new bone formation in group A was more abundant than that in group B, and it increased in group A but decreased in group B 3 months after implantation.The results confirmed that with higher BMD osseointegration was better. Osteoporotic changes could decrease OI and the amount of new bone formation at implant bone interface in rats. Osseointegration would not improve without outside factors.

Objective To study the value of serum galectin-3 detected by nanomagnetic beads sorting time resolved fluoroimmunoassay (TRFIA) tandem analysis in the diagnosis of pancreatic cancer.Methods The serum of 88 cases of pancreatic cancer,50 cases of acute pancreatitis,10 cases of pancreatic cysts,and 20 cases of healthy volunteers as control were collected.First,galectin-3 antibody coupled nanomagnetic-beads was used to sort the semm,then TRFIA was applied to detect the level of galectin 3,and the result was compared with that of routine TRFIA.ROC curve was constructed to determine the cutoff value and sensitivity for pancreatic cancer diagnosis.Results The method of nanomagnetic beads sorting TRFIA detected the level of galectin 3 between O.78 and 100 ng/ml in a linear manner,the intra-CV was ≤6.38％,and inter-CV was ≤7.22％,and average recovery rate was 105.3％.The serum level of galectin-3 in control group by nanomagnetic beads sorting TRFIA was 0.38 (0.02 ～ 3.06) ng/ml,which was significantly higher than that detected by routine TRFIA [0.18 (0.00 ～ 2.64) ng/ml,P =0.000).The serum levels of galectin-3 by nanomagnetic beads sorting TRFIA in pancreatic cancer,acute pancreatitis,pancreatic cysts and healthy volunteers were 4.85(0.00 ～42.67),0.69(0.00～ 13.62),0.70(0.00 ～ 14.54),0.38(0.02 ～ 3.06) ng/ml,and the level of galectin-3 in pancreatic cancer was significantly higher than that in acute pancreatitis,pancreatic cysts and healthy volunteers group (P ＜0.01),while the difference among the other three group was not significantly different.The AUC of galectin-3 for pancreatic cancer was 0.851 ± 0.040,95％ CI:0.772 ～ 0.929.While using 1.07 ng/ml as the cut-off value,the positive rates for the diagnosis of pancreatic cancer,acute pancreatitis,pancreatic cysts and healthy volunteers were 84.1％ (74/88),34.0％ (17/50),20.0％ (2/10),10.0％ (2/20),and the sensitivity for diagnosis of pancreatic cancer was significantly higher than those in other 3 groups (P ＜ 0.01).Conclusions Nanomagnetic beads sorting TRFIA tandem analysis method can enrich serum galectin-3,and increase the sensitivity of detection for pancreatic cancer and enhance the diagnostic value of galectin-3 for pancreatic cancer.

Objective To study the effects of asiaticoside on the growth of melanoma B16 cell cultures in vitro. Methods Melanoma B16 cells were subcultured and the inhibition of cellular growth was investigated. The morphology of the cells was observed after inhibition. The induction of apoptosis by asiaticoside was determined by flow cytometry. Results It was found that asiaticoside could significantly inhibit the growth of B16 cell cultures in vitro in a dose-dependent manner. The annexin-v positive cells were increased, along with that cells intaking R123 marked mitochondria were decreased, and PI positive cells increased, which indicated that cellular apoptosis was induced. Conclusion Asiaticoside plays an inhibitory role in the growth of melanoma B16 cells.

Objective To determine the risk factors for post-operative delirium(POD)and post-operative cognitive dysfunction(POCD)in patients undergoing spine surgery.Methods One hundred and twenty ASA Ⅰ-Ⅲ of both sexes aged 50-76 yr undergoing elective spine surgery under general anesthesia were studied.POD was assessed by Delirium Rating Scale revised 98 at 2 days after operation and the patients were assigned into POD and nonPOD group.Cognitive function was assessed by Mini-Mental State Examination(MMSE)at 1 day before and 3 days after operation.The patients were diagnosed as having POCD if MMSEpre-MMSEpost ≥ 3.The palients were assigned into POCD and nonPOCD group.Executive function and depression were assessed by stroop interference test and Beck Depression Inventory(BDI)at 1 day before operation.Age,sex,education,alcohol consumption per week,a history of psychiatric disease,ASA physical status,Charlson comorbidity score,type of anesthesia,anticholinergic drug administration and VAS score at 1 day after operation were recorded.If there was signifirant difference between the 2 groups,the factor was analyzed using multi-factor logistic regression to select risk factor for incidence of POD and POC).Results Eleven patients developed POD(9.2％)and 30 patients developed POCD(25.0％).Logistic regression model showed that lower Stroop-CW,higher BDI score,higher Charlson comorbidity score and a history of psychiatric disease were risk factors for POD,while lower Stroop-CW,higher BDI score,higher Charlson comorbidity score and higher alcohol consumption per week were risk factors for POCD.Conclusion Preoperative executive dysfunction,depression and greater preoperative comorbidity are risk factors for both POD and POCD.A history of psychiatric disease is a risk factor for POD and higher alcohol consumption is a risk factor for POCD in patients undergoing spine surgery.

Objective To investigate the effects of tg19320,a small peptide,interfering with IgG-FcγR interaction on the adhesion of neutrophils to endothelium and the expression of intercellular adhesion molecule 1 (ICAM-1)in endothelial cells and its possible mechanism.Methods Tg19320 was prepared by solid-phase peptide synthesis.ANCA IgG was isolated from the serum of active ANCA-associated systemic vasculitis(AASV)patients.When primary human umbilical vein endothelial cells(HUVEC)grew into connuence in cytokine-free eonditions,the cells were stimulated with TNF-α,human normal IgG,ANCA IgG and ANCA IgG+tg19320 respectively.HUVEC were pretreated with tg19320 for 45 minutes before being stimulated by ANCA IgG.Non-activated neutrophils was added to treat HUVEC and adhesion was measured by cell count.The expression of ICAM-1 mRNA and protein was assessed by real-time PCR and Western blot respectively.Soluble ICAM-1(sICAM-1)was determined using ELISA technique.Phosphorylation of IκB-α was assessed by Western blot. Results ANCA IgG significantly up-regulated the expression of ICAM-1 in HUVEC and promoted sICAM-1 release(P＜0.05),and TNF-α enhanced the effect of ANCA.These effects were almost completely abolished by tgl9320 both at protein and mRNA level.Furthermore,ANCA IgG increased the IκB-α phosporylation in HUVEC and tg19320could inhibit the effect. Conclusions ANCA IgG can modulate the expression of ICAM-1 and sICAM-1 release in endothelial cells.FcγR probably play a critical role in the ICAM-1 expression up-regulated by ANCA,which is mediated in part through NF-κB signaling pathway.Tg19320 has protective effect on endothelium in AASV in vitro.

Objective To study 6 type Ⅰ interferon (IFN)-inducible genes (IFIT4, IFI44, Ly6e,OAS1, OAS2 and OAS3) in patients with lupus nephritis (LN) and analyze its correlated expression levels with disease activity and/or clinical manifestations. Methods Total RNA was obtained simultaneously from kidney tissues and peripheral blood cells of 12 patients with diffuse proliferative lupus nephritis and 10 normal controls. Moreover, peripheral blood cells were obtained from 119 LN patients and 35 normal controls. Total RNA was extracted and reversely transcribed into complementary DNA. Gene expression levels were measured by real-time polymerase chain reaction by comparing to a housekeeping gene, and IFN score was calculated. Disease activity was determined by the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). Results The 6 genes were highly expressed in diffuse proliferative lupus nephritis patients compared with normal controls. IFN scores were positively correlated with SLEDAI score, the concurrent presences of anti-double-stranded DNA (anti-dsDNA) antibodies (P<0.05) and hypocomplementemia (P<0.01). Conclusion The 6 IFN-inducible genes are highly expressed iri LN patients. IFN scores are elevated in active lupus nephritis patients, in patients with positive anti-ds-DNA antibody and hypocomplementemia. IFN scores may be a useful biomarker for lupus nephritis therapy.