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 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

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.

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 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 explore the role of chemokines and ehemokine receptors in the etiopathog-enesis of diffuse proliferative lupus nephritis (LN). Methods ① Total RNA from the kidney tissues and peripheral blood cells of 12 patients with diffuse proliferative LN and 10 normal controls were prepared simultaneously and reverse transcribed into complementary DNA. Sybr green dye based real-time quantitative PCR method was used to compare the expression levels (indicated as-AACt value) of MCP-1, CCL19,CXCLg, CXCL10 and CCR2, CCR7, CXCR3. ② Immunofluoresceee labeling and immunohistochemical staining technique were used to observe the distribution of chemokines MCP-1, CCL19, CXCL9 and CXCL10 in normal and patients kidney tissues. Results The 4 chemokines genes (MCP-1, CCL19, CXCL9 and CXCL10) were consistently highly expressed in kidney tissues and peripheral blood ceils of diffuse proliferative LN patients compared with normal controls. The 2 chemokine receptors, CCR2 and CXCR3 were also overexpressed in peripheral blood cells of diffuse proliferative LN patients. There was nearly no expression of these 4 chemokine proteins in normal kidneys. But they were found in glomeruli of diffuse proliferative LN patients. Conclusion The expression of chemokines in the peripheral blood cells may be used as biomarkers for LN. Further study maybe lead to the development of specific drugs targeting at them for the treatment of systemic lupus erythematosus (SLE).

Objective To observe the expression of phosphoinositide 3-kinase(PI3K) and protein kinase B1 (Akt1) in PI3K/Akt signaling pathway in rat bones with fluorosis, and to reveal the mechanisms of the skeletal fluorosis. Methods Thirty-six SD rats were randomly divided into 3 groups (control group, low-dose fluorosis group, high-dose fluorosis group) and 12 rats were in each group according to body weight. The rats were fed with different concentrations of fluoride (NaF) to establish fluorosis models. Controls were fed with tap water( < 0.5 mg/L), experimental animals in low- or high-dose groups were fed with water containing NaF 5.0,50.0 mg/L, respectively. Rats were sacrificed after 6 months of treating with fluoride and the serum was kept for testing the bone metabolic markers of none gla protein(BGP) and cathepsin K(Cath-K) by enzyme-linked immunosorbent assay(ELISA), the proteins and mRNA levels of PI3K and Akt1 in rat bones were detected by immunohistochemistry and real time PCR, respectively. Results Each group of serum BGP and Cath-k were compared, the difference was statistically significant(F = 73.45,39.36, all P < 0.05). The contents of BGP[(1.99 ± 0.62), (2.38 ± 0.16)μg/L] and Cath-K [(89.07 ± 19.66), (110.16 ± 9.81)pmol/L] in the low-and high-dose fluorosis groups were higher than those in the control group[(0.15 ± 0.03)μg/L,( 18.32 ± 2.27)pmol/L], and the high fluorosis group was obviously higher than the low fluorosis group (all P < 0.05). Each group of serum PI3K and Akt1 protein and mRNA were compared, the difference was statistically significant(F- 178.16,118.08,38.81,52.31, all P< 0.05). Compared to the control group (181.55 ± 4.24,188.46 ± 2.18,3.84 ± 1.69,4.33 ± 0.89), the protein and mRNA expressions of PI3K(171.66 ± 2.85,154.12 ± 4.15,11.31 ± 4.18,20.54 ± 6.68), Akt1(177.47 ± 3.16,156.42 ± 3.18,12.52 ± 3.13,19.43 ± 5.36) were higher in the low- and high-dose fluorosis groups (all P < 0.05), and the high fluorosis group was obviously higher than the low fluorosis group (all P < 0.05). Conclusions BGP and Cath-K contents could be used as bone metabolic indices in the endemic fluorosis disease. Fluoride can increase the expression of PI3K and Akt1 mRNA and protein in bone tissue of fluorosis rats, and PI3K/Akt1 signaling pathway may be involved in the pathogenesis of bone injury caused by fluoride.