Objective:To investigate the effect of small interfering RNA (siRNA) silencing nicotinamide phosphoribosyltransferase (NAMPT) gene expression on proliferation and apoptosis of human multiple myeloma U266 cells and its mechanism.Methods:In vitro, NAMPT gene-specific siRNA was synthesized to transfect U266 cells. The experiment was divided into si-NAMPT group (transfected siRNA-NAMPT U266 cells) and si-NC group (transfected negative control siRNA U266 cells). The proliferation of U266 cells after transfection was detected by the methyl thiazolyl tetrazolium (MTT) method, and the apoptosis was detected by flow cytometry. Western blot was used to detect the expression levels of NAMPT, protein kinase B (AKT), phosphorylation-AKT (p-AKT), glycogen synthase kinase-3β (GSK-3β), phosphorylation-GSK-3β (p-GSK-3β), and β-catenin proteins in each group after transfection.Results:Compared with the si-NC group, the proliferation inhibition of U266 cells (absorbance value at 570 nm) increased after transfection for 48 h and 72 h in the si-NAMPT group (48 h: 0.78±0.06 vs. 1.62±0.11; 72 h: 1.23±0.14 vs. 2.37±0.18), and the differences were statistically significant ( t = 3.54, P = 0.034; t = 4.72, P < 0.01). The early apoptotic rate of cells in the si-NAMPT group increased compared with si-NC group [(53.42±0.25)% vs. (25.98±3.18)%], and the difference was statistically significant ( t = 4.41, P < 0.01). Compared with the si-NC group, the levels of p-AKT, p-GSK-3β, NAMPT and β-catenin proteins were significantly reduced in the si-NAMPT group (all P < 0.05). Conclusion:Silence of NAMPT gene can significantly inhibit U266 cell proliferation and induce cell apoptosis, and it may play a role by inhibiting the AKT-GSK-3β-β-catenin signaling pathway.

Objective To investigate the relationship between serum Leptin levels and insulin resistance.Methods During 2001-10~2002-08 fasting serum Leptin concentrations were measured with RIA in 196 cases(including 118 type 2 diabetes,78 controls),and height,body weight were measured to calculate body mass index(BMI).Results HbA_1c,SBP,TG,Leptin were correlated with ISI,Leptin concentration were higher in type 2 diabetes than in controls;Leptin concentration was significantly increased in obese subjects,whereas in type 2 diabetes serum Leptin concentration had no significant difference between obese subjects and non-obese subjects.Conclusion Most of obeses subjects have insulin resistance,whether obesity or not,the patients type 2 diabetes have some kind of insulin resistance.

Objective To explore MRI features of spinal canal enterogenous cysts.Methods The MRI features and differential diagnosis of 5 patients with spinal canal enterogenous cysts proved by surgery and pathology were reviewed retrospectively in combination with literature review.Results Of 5 cases,the cysts localized at cervical spine in 2,lumbar spine in 1,lumbosacral spine in 1 and the end of coccyx between rectum and cuticulum in 1.4 cysts were located at subdura,including anterior to the spinal cord in 3 and posterior to the spinal cord in 1.1 cyst located the end of coccyx between rectum and cuticulum was uncommunicated with spinal canal and 1 case associated with diastematomyelia.The spinal cords were compressed and displaced.The cysts were iso or slightly hyperintensity compared to CSF on T1WI,and similar intensity to CSF on T2WI.The cysts had no markedly enhancement on contrast-enhanced MR scan.Conclusion MRI has important value in diagnosing spinal enterogenous cysts.

Objective To investigate the effects of pretreatment with remifentanil on hemorrhagic shock (HS)-induced acute liver injury in rabbits.Methods Thirty-two New Zealand white rabbits,weighing 2.0-2.5 kg,were randomly divided into 4 groups (n=8 each): sham operation group (group S) ; group HS,low-dose remifentanil group (group RL) ; high-dose remifentanil group (group RH).Remifentanil was infused at 0.66 and 1.32 μg· kg-1 · min-1 for 145 min in groups RL and RH respectively,while the equal volume of normal saline was infused in group C.HS was induced by withdrawing blood from the left femoral artery at 15 min after continuous infusion of normal saline or remifentanil and mean arterial pressure (MAP) was reduced to 40 mm Hg.MAP was reduced to 35-45 mm Hg and maintained at this level for 120 min in groups HS,RL and RH.Blood samples were taken for determination of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities before administration (T0),immediately before blood letting (T1),and at 10,30,60 and 90 min after blood letting (T3-6).The animals were then sacrificed and the livers were immediately removed for microscopic examination.Results Compared with group S,the serum ALT activity at T5 and T6 and serum AST activity at T4-T6 were significantly increased in the other three groups (P ＜ 0.05).Compared with group HS,the serum ALT activity at T5 and T6,and serum AST activity at T4-T6 were significantly decreased in group RH,and no significant change was found in group RL (P ＞ 0.05).Compared with group RL,the serum ALT activity at T5 and T6,and serum AST activity at T4-T6 were significantly decreased in group RH (P ＜ 0.05).The serum ALT activity at T5 and T6 and serum AST activity at T4-T6 were significantly increased in groups HS,R and RH than those at T0 (P ＜ 0.05 or 0.01).The patho1ogical injury was attenuated in group RH compared with groups HS and RL.Conclusion Remifentanil pretreatment can attenuate HS-induced acute liver injury in rabbits,and the effect is related to the dose.

Objective:To evaluate the effects of different denture cleaners on the discoloration of heat-cured denture base resin and artificial teeth.Methods:40 same specifications of the heat-curing denture base resin and 40 artificial central incisors were immerced in an acombination stain of coffee,tea and soy sauce for 4 weeks.Then specimens and artificial teeth were randomly distributed into 4 groups and soaked in Polident,Steradent,0.5％ sodium hypochlorite solution and distilled water for 4 hours respectivelly(n =10).Color differences(△E) were measured by using a colorimeter and a denture spectrophotometer before and after staining,and after cleaning.Results:Before or after staining there was no difference of △E among the groups of denture base risn or artificial teeth(P ＞ 0.05).After cleaning the denture base resin and the artificial teeth in the group of 0.5％ sodium hypochlorite solution presented higher △E than the other groups(P ＜ 0.05),in group of distilled water presented lower △E than the other groups(P ＜ 0.05).No significant difference was found between Polident group and Steradent group(P ＞0.05).The △E of the denture base resin and artificial teeth.in distilled water group before staining and after cleaning were the highest among the groups(P ＜0.05).Conclusion:0.5％ sodium hypochlorite solution,Polident and Steradent are effective in removing the discoloration from the heat-cured acrylic resin and artificial teeth.0.5％ sodium hypochlorite solution is the most effective,Polident and Steradent are the similar.

Objective To explore the distribution and antimicrobial resistance of pathogens causing healthcare-associated infection(HAI) in a hospital,and provide theoretical basis for rational use of antimicrobial agents and control of HAL Methods HAI occurred in patients in this hospital between 2011 and 2015 were chosen,distribution of major pathogens causing HAI and antimicrobial resistance were analyzed.Results A total of 852 pathogenic strains were isolated,including 458 (53.75 ％) strains of gram-negative bacteria,259 (30.40 ％) fungi,and 135 (15.85 ％) gram-positive bacteria,the top 4 pathogens were Klebsiella pneumoniae (n =159,18.66 ％),Candida albicans (n =119,13.97％),Acinetobacter baumannii (n =81,9.51％),and Escherichia coli (n =74,8.69 ％).Isolation rate of gram-negative bacteria increased from 47.41 ％ in 2012 to 61.57％ in 2015,fungi increased from 24.78％ in 2011 to 34.45％ in 2014 (2015 was 27.95％),gram-positive bacteria decreased from 26.55％ in 2011 to 10.48％ in 2015.Resistance rates of Escherichia coli and Klebsiella pneumoniae to imipenem and meropenem were 8.11％-12.58％;Acinetobacter baumannii had the lowest resistance rate to cefoperazone/sulbactam(23.46％),resistance rate of Candida albicans to fluconazole was up to 59.66％,vancomycin-resistant gram-positive bacteria was not found.Conclusion The major pathogens causing HAI are opportunistic pathogens,antimicrobial resistance is serious,the species and distribution varies with different years,it is necessary to enhance specimen detection rate,strengthen the monitoring on pathogen resistance,and use antimicrobial agents rationally.

Objective To investigate the effects of high-fat diet on pancreatic acinar cells' IP3 expression and CCK-induced amylase release in rats.Methods Male Wistar rats were divided into high-fat diet group and normal diet group,they were fed for 4 weeks.Blood triglycerides,cholesterol,amylase and glucose levels were determined by automatic biochemical analyzer.Pancreatic tissues were taken for histopathological observations.Pancreatic acinar cells were isolated and cultured,and intracellular inositol 1,4,5-trisphosphate (IP3) was detected using a commercial kit.Amylase release rates were measured after CCK-8 stimulation.Results The rats in high-fat diet group appeared hyperlipidemia,vacuolization of acinar cells and the lymphocytes appeared around the acinar cells can be seen on the pancreatic tissue pathology staining.The levels of IP3 in acinar cells of rats fed a high-fat diet were higher than that of normal rats [(31.807 ± 3.448) pmol/106 cells vs (24.632 ± 3.649) pmol/106 cells,t=7.479,P＜0.001];and amylase release rate in these rats'acinar cells were also higher than those of normal rats [when CCK-8=0.01 nmol/L:( 11.056 ±3.369)％ vs (7.354 ± 2.181) ％,t=3.912,P＜0.001;when CCK-8=1 nmol/L:( 13.854 ± 4.087 ) ％ vs (9.432 ±2.477) ％,t=3.939,P＜0.001 ) after CCK-8 stimulation in different concentrations.Additionally,there was a positive co-relationship between acinar cell's IP3 level and amylase release (r=0.896,P＜0.001 ).Conclusion Chronic high-fat diet induces hypersensitivity for pancreatic acinar cells' exocrine function,and IP3 as a signal molecule may play an important role in this process.

BACKGROUND:Preliminary data showed that the application of human platelet lysate to human umbilical cord mesenchymal stem cell culture can better maintain the characteristics of stem cells than the application of serum-free medium. However, the serum-free medium can better improve the proliferation of mesenchymal stem cells in vitro than the human platelet lysate.OBJECTIVE:To screen out a better mesenchymal stem cell cultivation system that can greatly maintain the characteristics and proliferation rate of human umbilical cord mesenchymal stem cells.METHODS:Six human umbilical cord specimens were inoculated in six culture systems, and the primary culture of umbilical cord mesenchymal stem cell was performed. These six culture systems were respectively MesenCult serum-free medium with 2% human platelet lysate (group A), StemPro serum-free medium with 2% human platelet lysate (group B), MesenCult serum-free medium (group C), StemPro serum-free medium (group D), low glucose-DMEM with 10% fetal bovine serum (group E), low glucose-DMEM with 10% human platelet lysate (group F). The cells were subcultured at 14 days after inoculation to compare the effects of different culture systems on the morphology, surface markers, differentiation and proliferation of human umbilical cord mesenchymal stem cells.RESULTS AND CONCLUSION:(1) The morphology of passage 3 cells in group D was elongated and uneven in size. The morphology of passage 3 cells was flattened in groups E and F, but the cells in the other groups were spindle-shaped and uniform. There were no significant changes in morphology and size between passage 3 and 5 cells in A and B. In group C, the morphology of passage 5 cells was more flattened and uneven in size compared with passage 3 cells. In group D, the morphology of passage 5 cells was more elongated than that of passage 3 cell. In group E, the morphology of passage 5 cells was more flattened than that of passage 3 cells. There was no significant difference in morphology between passage 3 and 5 cells in group F. (2) The expression rate of cell surface markers had no significant difference at different passages in each group. (3) The adipoinduction and osteoinduction rates were relatively higher in groups A and B compared with groups E and F, and lowest in groups C and D. (4) The cell proliferation rate for each passages in group A was significantly higher than that in group C. The cell proliferation rate for each passage in group B was significantly higher than that in group D. The cell proliferation rate for each passage in groups E and F was significantly lower than that in groups A and B. To conclude, these results suggest that the combination of serum-free medium with human platelet lysate could better maintain the characteristics and the proliferation efficiency of mesenchymal stem cells.

Objective To study direct economic loss of healthcare-associated infection (HAI)in hospitalized children with acute respiratory infection (ARI),and the influence in satisfaction degree of children’relatives.Methods Clinical data of 1 039 children with ARI in a pediatrics ward between April 1,2012 to March 31,2013 were analyzed retrospective-ly.50 patients with HAI were in case group and 50 patients without HAI were in control group.The difference in medical cost and the degree of satisfaction of children’s relatives were compared between two groups.Results Medical cost of pa-tients in case group was significantly higher than control group([P 25 - P 75 :￥3 095.54 - ￥4 779.48]vs [P 25 -P 75 :￥1 744.14-￥2 382.07],Z =5.89,P <0.01).The expenses in medicine,laboratory examination,diagnosis and treatment,and nursing in case group were all higher than control group,there were significant differences between two groups(all P <0.01 );Hospitalization day of case group was significantly longer than control group ([P 25 -P 75 :9 d-15 d]vs[P 25 -P 75 :6 d-8 d],Z =5.79,P <0.01 ).Satisfaction degree of patients’relatives in case group was significantly lower than control group (Z =8.22,P <0.01).Conclusion HAI occurred in ARI children can increase medical cost,prolong length of hospital stay and decrease satisfaction degree of patients’relatives.

Objective:To study the correlation of β-fibrinogen-854G/A,-455G/A,-249C/T,-148C/T,448G/A and BcI-1G/A polymorphisms, functional expression of plasma fibrinogen concentration, molecular reactivity, and the type of cerebral infarction.Methods: A casecontrol study was used to analyze 54 patients with main-think cerebral infarction(MCI), 106 patients with penetrating-arterial cerebral infarction (PCI)and 160 heathy cases as control group in Kailuan Hospital between July 2002 and June 2003.Gene polymorphisms were detected by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP).Fg concentration, fibrin monomer polymerized velocity(FMPV), absorbance maximum(Amax), FMPV/Amax and biochemistry factors including TG were measured, Results: Fg concentration, FMPV, FMPV/Amax in the MCI group and TG, VLDL and FMPV in the PCI group were higher than in the control group(P＜0.05).The frequencies of854A and Bcl-1A alleles had significant difference among three groups,and the frequencies of GA and AA genotypes in the MCI and PCI groups were higher than in the control group(P＜0.05), however, no different genotypes and allele frequencies of the remaining sites were found in the three groups(P＞0.05).Fg concentration and FMPV of allele T carriers in the MCI group were less than that of-249C/C homozygous ones(P ＜0.05); FMPV/Amax of allele T carriers in the PCI group was higher than that of-148C/C homozygous ones(P＜0.05);with allele A carriers, Fg concentration of control group and FMPV of PCI group were higher than that of Bcl-1 wild homozygote(P＜0.05).Conclusion: Bβ-249 C/T polymorphism in the 5-flanking promoter region can influence the expression of plasma FMPV, Bβ-148 locus is the main regulation location of Fg molecular conglomerate function.Bcl-1 locus in the 3-flanking region is an important gene regulator of plasma Fg concentration, moreover,people with its mutated genotypes are susceptible to MCI.The abnormal plasma Fg concentration, FMPV/Amax and FMPV simultaneously are important risk factors for MCI, and only abnormal FMPV and TG are prone to PCI.