Objective Thymidine phosphorylase (TP) cDNA was transfected into colorectal cancer cell lines LOVO with the lentiviral vector,the anticancer effeciency of 5'-deoxy-5-fluorouridine (5'-DFUR)and 5-fluorouracil (5-FU) on LOVO cells were evaluated.Methods TP cDNA were transfected into LOVO cell line with the lentiviral vector pLenti6.3_MCS_IRES2-EGFP,and the transfection efficiency was analyzed by flow cytometer and immunohistochemistry.Cells were divided into six groups:LOVO,LOVO-TP,LOVO-control,LOVO + INF-α2a,LOVO-TP + INF-α2a,LOVO-control + INF-α2a.TP protein expression and the relative quantitative analysis for TP mRNA in transfections cells were detected by Western blot and RT-PCR respectively.Volumes of converted 5-FU from either in the medium containing different concentration of 5'-DFUR,in which all cells were cultured,or in cells lysate,were detected by high performance liquid chromatography (HPLC).Results The transfection efficiency of TP cDNA in LOVO cells was 95％.The Mean gray value of TP protein expression in LOVO-TP and LOVO-TP + INF-α2a were 198.15 folds and 243.22 folds higher than LOVO cell,respectively (P ＜ 0.01).The RQ values of TP mRNA expression in LOVO-TP and LOVO-TP + INF-α2a were also 18.56 folds and 59.61 folds higher than wild LOVO cell,respectively (P ＜ 0.01).The IC50 values of 5'-DFUR on LOVO-TP and LOVO-TP + INF-α2a were 1 660 μ mol/L and 813 μ mol/L,respectively,significantly lower than 4 462.59 μ mol/L in wild LOVO (P ＜ O.01).The 5-FU volumes detected from media contained series concentration of 5'-DFUR for culturing LOVO-TP and LOVO-TP + INF-α2a were 2.0-5.3 folds,and 2.9-10.4 folds more than wild LOVO,respectively (P ＜ 0.01).Conclusions Transfected TP cDNA into colorectal cancer cell line LOVO with lentiviral vector increases the expression of TP mRNA and TP protein and the amount of 5-FU converted from 5'-DFUR,enhancing its anticancer effect.

Objective To evaluate the capabilities of disc diffusion and Vitek2-compact GN13 methods for testing antimicrobial susceptibility of screening ESBLs ( extended-spectrumβ-lactamase) in En-terobacteriaceae clinical isolates.Methods A total of 93 Enterobacteriaceae strains were isolated from pa-tients with intra-abdominal infections in 21 hospitals during 2011 to 2012.The in vitro minimum inhibition concentration ( MIC ) values of ampicillin-sulbactam, piperacillin-tazobactam, ertapenem, ceftazidime, ceftriaxone, cefepime, imipenem, amikacin, ciprofloxacin and levofloxacin were determined by disc diffu-sion, Vitek2-compact GN13 and broth microdilution methods, respectively.Categorical agreement ( CA ) rates of disc diffusion and Vitek2-compact GN13 methods were determined by using broth microdilution meth-od as the reference method.The genes encoding ESBLs were screened in Escherichia coli (E.coli), Kleb-siella pneumoniae (K.pneumonia), Klebsiella oxytoca (K.oxytoca) and Proteus mirabilis (P.mirabilis) strains by using PCR analysis and gene sequencing.Disc diffusion and Vitek2-compact GN13 methods were used for the phenotypic confirmatory test of ESBLs and the sensitivity, specificity, positive predictive value and negative predictive value of the two tests were evaluated.Results The CA values of disc diffusion and Vitek2-compact GN13 methods for the 10 antibiotics were all >90% as compared with broth microdilution method.The major error (ME) rate for ertapenem was 3.2%and the very major error (VME) rates for am- picillin-sulbactam, ceftazidime and cefepime tests were all 2.2% by using Vitek2-compact GN13 method. The sensitivity, specificity, positive predictive value and negative predictive value of disc diffusion and Vitek2-compact GN13 methods in the phenotypic confirmatory test of ESBLs were 96.7%(29/30), 100%(20/20), 100%(30/30) and 95%(19/20), respectively.Conclusion Both disc diffusion and Vitek2-compact GN13 methods could be used for testing the antimicrobial susceptibility and the detection of ESBLs in Enterobacteriaceae clinical isolates with the advantage of accuarcy.Attention should be paid to the posibil-lity of oaurance of ME and VME when testing ertapenem, ampicillin-sulbactam, ceftazidime and cefepime by using Vitek2-compact GN13 method.

Background and purpose:Life expectancy has continuously improved in recently years with the development of medical level. In 2012, the amount of people who were above 60 years old has reached 810 million and account for 11% of worldwide population. The worldwide population shift towards older ages will inevitably lead to more elderly patients being diagnosed with non-small cell lung cancer (NSCLC). It still remains controversial whether sublobar resection is effective in such cases. In order to solve this question, we need to understand the clinical characteristics of these tumors. Methods:From 2006 to 2012, a total of 310 patients with NSCLC who were above 65 years old underwent surgical resection in Department of Thoracic Surgery, the First Afifliated Hospital of China Medical University and the Second Department of Thoracic Surgical Oncology, Jilin Province Tumor Hospital. The clinical data were retrospectively analyzed in sex, stage, histology, smoking status, smoking amount, drinking status, surgical approaches, multimodality therapy and overall survival. Survival was analyzed by Kaplan–Meier method and log-rank test. Results:There were 256 (82.6%) elderly patients who underwent standard lobectomies and 54 (17.4%) patients who underwent sublobar resections. The overall 5-year survival rate was 52.9%. Patients with different surgical approaches (lobectomy and sublobar resection) had nearly the same 5-year survival rates (60.5%vs 60.8%, P=0.381). The prognosis were signiifcantly inlfuenced by gender (P=0.024), stage of disease (P=0.028), smoking status (P=0.034) and smoking amount (P=0.028). The multivariate Cox proportional hazards analysis revealed that the smoking status was associated with the hazard ratio of 1.660 (1.058-2.606;P=0.028). Conclusion:In elderly NSCLC patients, sublobar resection is considered to be an appropriate treatment in comparison with lobectomy, as this procedure provides an equivalent long-term survival. The survival of elderly patients with lung cancer is closely related to the smoking status.

Objective To study effect of rubescens on inflammatory cytokines and liver function of alcoholic liver injury rat model.Methods 120 SD rats were chosed for studied, randomly divided into control group, model group and treatment group,40 rats per group.Alcoholic liver injury rat model was made by ethanol feeding approach, rubescens were given for treatment.Then serum AST, ALT, α-GST, GLDH content and liver tissue NF-κB, TNF-α, IL-6, IL-8 levels were detected by electrochemiluminescence and fluorescent quantitation PCR,respectively.Results Model group serum AST, ALT,α-GST, GLDH contents and NF-κB, TNF-α, IL-6, IL-8 contents in liver tissue were higher than control group(t=12.030~37.417, P<0.05). Treatment group serum AST,ALT, α-GST, GLDH contents and NF-κB, TNF-α, IL-6, IL-8 contents in liver tissue were lower than model group (t=5.128~32.325, P<0.05).Conclusion Alcohol feeding can cause liver damage, rubescens therapy can reduce liver injury, relieve inflammation.

Objective To study the role of serum chemokines and their receptors in the pathogenesis of atopic dermatitis(AD).Methods Serum levels of interferon ?-inducible protein-10(IP-10),stromal cell-derived factor-1? (SDF-1?),eotaxin,thymus and activation-regulated chemokine(TARC) as well as macrophage-derived chemokine (MDC) were detected by enzyme-linked immunosor bent assay(ELISA) in 39 patients with AD and 39 normal controls.Meanwhile,chem okine receptors CXCR3,CXCR4,CCR3,CCR4 and CCR5 on peripheral blood CD4+T-lymp hocytes were analyzed by flow cytometry with a two-color immunofluorescent sta ining in 39 patients with AD.Results Serum levels of SDF-1?,TARC and MDC we re significantly higher(P 0.05) between the patients[(123.6 ? 110.4) pg/mL a nd (68.7 ? 26.2) pg/mL,respectively]and controls [(100.7 ? 73.7) pg/mL and(66.8 ? 20.5) pg/mL,respectively].The expression of CXCR3,CCR3,CCR4 and CCR5 o n CD4+T cells was increased (P

Objective To evaluate the characteristics of the PREVI Isola automated plate streaker (bioMérieux,SA).Methods 80 respiratory tract specimens,70 sterile fluids,52 stools,69 swabs,12 cerebrospinal fluids and 80 urines were collected in Peking Union Medicd College Hospital.Specimens were processed with manual streaking and PREVI Isola system.PREVI Isola system were evaluated comparing to the manual streaking.The quality of results were analyzed by SPSS 16.0, doing Wilcoxon’s Sign Rank Test for the results of finally isolated species,overall numbers of isolated colonies and semi-quantita-tive of the species were both isolated by the two methods.Results PREVI Isola system was highly automatic,which could select the right plates and stake the bar code on the back of the plate indicating the type of the agar and inoculation time, PREVI Isola system could accurately absorb the liquid specimen and use a novel comb streaking procedure for processing of fluid specimens on standard agar plates,like 17 inoculating loops work together.It also had a good reproducibility.The quali-ty of PREVI Isola system results:As to the finally isolated species,there was significant statistical difference between PREVI Isola system and manual streaking method in respiratory tract and stools specimen,there were more species isolated by manual streaking method than PREVI Isola system.There were no differences between the two methods for the other of specimen types.As to the amount of pure clones of the species were both isolated by the two methods,there were significant statistical differences between the two methods for respiratory tract,sterile fluid and stool specimens.The amount of clones isolated by PREVI Isola system was more than manual streaking method.In semi-quantitative results,there were significant statistical differences between the two methods for respiratory tract and urine specimen,Species had wider distribution of PREVI Isola system than manual streaking method.Inoculation efficiency:if the batch of specimen type was simple (mainly the urine and so on),using the same plates,PREVI Isola system was more efficient than manual streaking method.Howev-er,if the batch of specimen type was complicated,manual method was high-performance.Besides,not all specimen type could be inoculated by PREVI Isola system,such as cerebrospinal fluid,catheter and tissues.Conclusion If the lab had simple specimen type,or utilize the specimen type using the same agar plates to be inoculated together,PREVI Isola system belongs to a good performance automated plate streaker.

OBJECTIVETo explore the action mechanism of electroacupuncture (EA) at "Shenmen" (HT 7) and "Sanyinjiao" (SP 6) on insomnia.

METHODSThirty SD rats were randomly divided into a blank group, a model group and an EA group, 10 cases in each group. The insomnia model was made by immobilization method in the model group and EA group. After model establishment, rats in the blank group and model group were treated with fixation and no treatment was given. Rats in the EA group were treated with EA at "Shenmen" (HT 7) and "San-yinjiao" (SP 6) for 15 min, once a day for 4 days. After treatment, the level of daytime and nighttime activity, open-arm entry percentage and open-arm time percentage of elevated plus-maze test were measured; the content of noradrenaline (NE), dopamine (DA) and epinephrine (EPI) in plasma and NE, DA in thalamus and brainstem were detected by using euzymelinked immunosorbent assay method.

RESULTSCompared with the blank group, the daytime activity was increased and nighttime activity was reduced in the model group (both P<0. 05); the open-arm entry percentage and open-arm time percentage of elevated plus-maze test were both reduced in the model group (both P<0. 05); the contents of NE, DA, EPI in the plasma and NE, DA in thalamus and brainstem were increased in the model group (all P<0. 05). Compared with the model group, the daytime activity was reduced and nighttime activity was increased in the EA group (both P<0. 05); the open-arm entry percentage and open-arm time percentage of elevated plus-maze test were both increased in the EA group (both P<0. 05); the contents of NE, DA, EPI in the plasma and NE, DA in thalamus and brainstem were reduced in the EA group (all P< 0. 05).

CONCLUSIONElectroacupuncture at "Shenmen" (HT 7) and "Sanyinjiao" (SP 6) can restrain the over-activity of the sympathetic-adrenal medullary system to treat the insomnia.

Acupuncture Points ; Adrenal Medulla ; metabolism ; Animals ; Anxiety ; Disease Models, Animal ; Dopamine ; metabolism ; Electroacupuncture ; Female ; Humans ; Male ; Norepinephrine ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sleep Initiation and Maintenance Disorders ; metabolism ; physiopathology ; psychology ; therapy ; Sympathetic Nervous System ; physiopathology

Objective To investigate the effect of curcumin on Aβ production in swAPP HEK293 cells and its preliminary mechanism.Methods swAPP HEK293 was used as cell model,the effect of curcumin (at different time points and of concentration) on cell viability was accessed by MTT assay.After the cells were treated with non-cytotoxic concentration of 5 μmol/L for different time,ELISA was used to detect Aβ production.The concentration and the time point of the strongest inhibitory effect were selected for the following tests.Real time PCR was employed to analyze miR-153 and APP mRNA expression,and Western blot was used to detect APP protein expression.Results As compared with the control group,curcumin of ≤ 5 μmol/L had no toxicity effect on the cell viability (P ＞ 0.05).Curcumin significantly inhibited Aβ production (P ＜ 0.05).Therefore 5 μmol/L curcumin and 24 h were selected as the best concentration and timc.Curcumin of 5 μmol/L had no obvious impact on APP mRNA expression (P ＜ 0.05),whereas markedly decreased APP protein expression.In addition,miR-153 level in the cells was significantly increased by 5 μmol/L curcumin treatment (P ＜ 0.05).Conclusion Curcumin may inhibit Aβ production through up-rcgulating miR-153 level and reducing APP protein expression in swAPP HEK293 cells.

BACKGROUND:Whether plumbum(Pb) exposure induced learning and memory disorder in animals is correlated with the changes of the activity of protein kinase C(PKC) in the brain tissue? OBJECTIVE:To investigate the alterative rules of PKC activity in the brain tissue of rat in development stage under chronic exposure of Pb and its effects on the memory development of the mice. DESIGN:A randomized controlled study employing experimental animals as subjects. SETTING: Cellbiology Key Laboratory of the Ministry of Health,China Medical University. MATERIALS:The study was conducted in the Laboratory of Biochemistry and Molecular Biology of China Medical University. Totally 72,5- 6 weeks old Kunming mice were selected. METHODS:Lead acetate of different concentration was used to feed female mice after copulation.Fetal mice contacted Pb through breast feeding and drinking water.Fetal mice were executed separately on 1st day(P1),8th day(P8),15th day(P15),22nd day(P22), and 30th day(P30) after birth for brain tissue harvesting.PCK activity in the brain tissue of the mice exposed to Pb was detected in vitro by[γ 32P] ATP.Budding mice were exposed to lead acetate of different concentration for the observation of the effects of Pb in difference concentration on the memory of the mice through memory behavioral training and test by passive avoidance response experiments. RESULTS:PKC activity detection indicated that PKC activity in brain of Pb exposed budding mice during initial growth stage was higher than normal,while it was lower than normal during advanced stage of growth.High concentration Pb had relative stronger inhibition on PKC activity.Memory behavioral training indicated that low concentration Pb would induce elevation of memory curve in the initial stage of the development in mice but reduction of memory curve in middle and long term.The increase of Pb concentration would reduce memory curve. CONCLUSION:Pb has inhibitive effects on the development of PKC activity in brain tissue of mice:the higher the Pb concentration is, the more significant the inhibition is.Low concentration Pb seems to have stimulatory effects on memory in short period while long term exposure to Pb would induce memory inhibition.The inhibitive effect of high concentration Pb is more significant.Pb has certain correlation with PKC activity in brain tissues of budding mice and memory function.

Objective To evaluate antimicrobial resistance and antimicrobial resistance mechanisms of Enterococcus faecalis (E.faecalis)to linezolid (LNZ),and provide basis for clinical rational drug use.Methods Twelve E.faecalis strains isolated from sputum of patients who received LNZ therapy in a hospital between January 2012 and January 2013 were collected.The minimum inhibitory concentrations (MICs)of antimicrobial agents were de-termined by agar dilution method,23S rRNA V region gene of E.faecalis was amplified by polymerase chain reac-tion,the amplified products were sequenced.Results Of 1 2 isolates,2 were intermediate strains and 1 0 sensitive strains.The G2576U mutation was detected in 2 intermediate strains,1 of which was also detected G2424U muta-tion;the variations were not detected in 10 sensitive strains.C2424U and G2576U mutation existed in R1 and R4 region respectively.Conclusion 23S rRNA V region gene mutations are found in the intermediate strains of E.faecalis.Change in MIC values of linezolid should be paid close attention in clinical use.