OBJECTIVE Topreparealipidderivativeofgemcitabine(Gem)anditspolymericmi-celles to overcome the disadvantages of Gem.METHODS N-benzyl-3′-acetyl-gemcitabine(BAG)was synthesized.A BAG-loaded poloxamer polymeric micelle (BAG∶poloxamer 188 =10∶1 ,mol/mol)was prepared using an injection method.The micelles were characterized with a laser particle size and elec-tric charge instru ment and negatively-stained trans mission electron microscopy.Hu man breast cancer cells MCF-7 were cultured with Gem or BAG polymeric micelles of 5,10,20,30,50,70,90 μmol·L-1 for 24,48 and 72 h,respectively.The inhibitory rate of cells was measured with an MTT method.The MCF-7 cytotoxicity of BAG polymeric micelles was investigated.A pharmacodynamic study was per-formed on the mice bearing mouse hepatocellular cancer cells H22.Intravenous (iv)and oral (ig)ad-ministration was used at the dose of Ge m 40 mg·kg -1 or BAG polymeric micelles 62 mg·kg -1 .The mice were administered on the 1 st,4th and 7th day and sacrificed on the 8th day.Tumor inhibitory rates were measured.RESULTS TheBAGstructurewasidentifiedbythinlayerchromatograph,1Hand13C NMR,infrared ray chromatograph and mass spectrum.The appearance of BAG micelles was a slightly blue suspension.The micelles were spheres according to the electron microscopic observation.Their size was 62.82 nm and the zeta potential was -18.8 mV.The half inhibition concentration (IC50)of Gem and BAG polymeric micelles was 40.6 and 90.0 μmol·L-1 ,5.0 and 14.9 μmol·L-1 ,5.0 and 1 3.6 μmol·L-1 at 24,48 and 72 h,respectively according to the MTT results.According to the in vivo results,compared with the tumor model group,Gem (ig),Gem (iv)and BAG polymeric micelles (iv and ig)had significant effect on the tumor weight of H22 cell xenograft mice (P<0.01 ).As for anti-tumor efficiency,BAG polymeric micelles (ig)were better than Gem (ig)(P<0.05);BAG polymeric micelles (iv)were better than BAG polymeric micelles (ig)(P<0.05),and BAG polymeric micelles (iv)were almostequaltoGem(iv).CONCLUSION ThelipidderivativeofGemcanbeloadedinthepoloxamer 1 88 polymeric micelles.BAG polymeric micelles show in vitro MCF-7 cell inhibition and in vivo inhibition of mouse H22 xerografts;iv or ig.BAG polymeric micelles (ig)show better anti-tumor effect than Gem (ig),indicating that BAG polymeric micelles are a promising novel anti-tumor oral preparation.

Objective To investigate the therapy and influencing factors for prognosis of ventilator-associated pneumonia(VAP) caused by multidrug-resistant organisms(MDROs).Methods 169 patients with VAP who were admitted to a hospital between January 2012 and December 2013 were included in analysis, 125 were in MDRO infection group and 44 in non-MDRO infection group.MDRO infection group was subdivided into MDR-A group(n=78, resistant to selected antimicrobial agents) and MDR-B group (n=47, sensitive to at least one kind of selected antimicrobial agent).Antimicrobial choice and prognosis between each group were analyzed and compared.Results 242 strains of pathogenic bacteria were isolated from airway secretion of VAP patients, 173(71.49％) were MDROs.The major pathogens causing VAP were Klebsiella spp.(n=66), Pseudomonas aeruginosa(n=64), Acinetobacter spp.(n=60), Staphylococcus aureus(n=27), and Escherichia coli (n=17), the percentages of MDROs of above pathogens were 68.18％, 50.00％, 91.67％, 88.89％, and 76.47％ respectively.The prognosis of MDRO infection group was poorer than that of non-MDRO infection group, MDR-A group had the worst prognosis(P<0.001).Persistent fever, leukocytosis, and progress of pulmonary inflammation in VAP patients suggested poor prognosis(all P<0.001);antimicrobial use in patients with effective therapy was higher than those in a worsened condition before onset, at the beginning of onset, and after culture of specimens(all P<0.001), while coma, early-onset VAP and multiple bacterial infection had no prognostic significance in patients with VAP(all P>0.05).Conclusion There is high incidence of MDRO infection in patients with VAP, effective antimicrobial therapy can improve the prognosis.

ObjectiveTo investigate the changes in the expression of T-cell death-associated gene 8(TD- AG8) in spinal cord in rats with bone cancer pain.MethodsTwo hundred and twenty-four female rats weighting 150-180 g were randomly divided into 3 groups:normal control group(group Ⅰ,n ＝ 64),normal saline group (group Ⅱ,n ＝ 64),bone cancer pain group(group Ⅲ],n ＝ 96).Bone cancer pain was induced by inoculating Walker256 mammary gland carcinoma cells into the tibia medullary cavity.Mechanical withdrawl threshold(MWT)was measured at 1 d before(baseline)and 1,3,6,9,12,15 and 18 d after inoculation.Sixteen rats were sacrificed at 1 day before(baseline)and 6,9,12,15 and 18 d after inoculation in group Ⅲ and 18 d after inoculation in groups Ⅰ and Ⅱ.The L4-6 spinal cord were removed,and the number of TDAG8 positive cell was counted,and the expression of TDAG8 mRNA was measured by RT-PCR.ResultsCompared with baseline value and group Ⅰ,MWT was decreased,and the number of TDAG8 positive cells and the expression of TDAG8 mRNA in spinal cord were increased at 6-18 d after inoculation in group Ⅲ ( P < 0.01 ).ConclusionThe expression of TDAG8 in spinal cord is up-regulated in rats with bone cancer pain,which may be involved in the mechanism of the development of bone cancer pain.

Aim To pick out the siRNA which could most effectively inhibit the expression of brain-derived neurotrophic factor( BDNF) in microglial cells,to detect the cytotoxicity of the transfection complex,and to ob-serve the change of OX-42 expression,the microglial marker,after BDNF siRNA treatment. Methods Four siRNAs were chemically synthesized: three of them were used to inhibit BDNF expression in microglial cells,the rest was fluorescence-labeled mismatch siRNA as a negative control. They were all transfected into microglial cells,respectively. BDNF mRNA was detected 24 h after transfection by Real-Time PCR and itsprotein expression was observed done by Western blot 48 h later. The Sulforhodamine B( SRB) assay was used to investigate the drug-induced cytotoxicity. Co-expres-sion pattern of BDNF and OX-42 was determined by double-labeling immunofluorescence. Results ① The BDNF siRNA1588 was the most effective siRNA,compared with the vehicle or mismatch siRNA-treated group( P

Objective To investigate the effect of video-assisted thoracoscopic surgery( VATS) in thoracic disease,and the feasibility to carry out VATS for basic hospital. Methods The data of VATS treatment were collected to compare the differences between study group and control group,and evaluate the the feasibility to carry out VATS for basic hospital. Results The operation time was (100. 75±22. 72) min, blood loss was (54. 27±26. 21) mL,postoperative drainage was (920. 67±171. 99) mL. The postoperative complications were fewer,post-operative hospital stay was shorter,incision time was shorter(P=0. 000) and pain scores was lower(P=0. 000) in study group than that in control group. Basic hospital has the capacity to conduct this technical. Conclusion VATS is feasible to carry out in basic hospital.

Objective To investigate the role of P2Y1 purinergic receptors in the spinal cord in a rat model of bone cancer pain. Methods Ninety female SD rats weighing 150-180 g were randomly divided into 5 groups (n = 18 each): Ⅰ group sham operation; Ⅱ group bone cancer pain; Ⅲ group sham operation + MRS2179 (a specific P2Y1 purinergic receptor antagonist); Ⅳ group BCP + vehicle (group NS); Ⅴ group BCP+ MRS2179.Bone cancer pain was induced by inoculating Walker 256 mammary gland carcinoma cells into medullary cavity of tibia. In group Ⅲ, Ⅳ, Ⅴ MRS2179 100 pmol/10 μl or NS 10 μl was injected intrathecally once a day for 3 days starting from the 7th day after operation. Mechanical pain threshold to von Frey stimuli was measured before and every other day after operation. The anirnals were sacrificed on the 9th day after operation. The L4-6 segment of the spinal cord was removed for detection of expression of P2Y1 receptor and phosphorylated extracellular signal-regulated protein kinase 1/2 (p-ERK1/2) in the spinal dorsal horn. Results P2Y1 receptors and p-ERK1/2 coexisted in spinal dorsal horn. Inoculation of cancer cells into tibia significantly decreased mechanical pain threshold at postoperative day 6-18 and increased the expression of P2Y1 receptor and p-ERK1/2 on the 9th day after operation in group Ⅱ and Ⅳ as compared with group Ⅰ . Intrathecal MRS 2179 significantly increased pain threshold and decreased expression of P2Y1 receptor and p-ERK1/2 in group Ⅴ compared with group Ⅱ and Ⅳ. Conclusion P2Y1 receptors in the spinal cord are involved in the development of bone cancer pain, which may be related to the activation of ERK1/2.

Objective To evaluate the role of brain-derived neurotrophic factor (BDNF) in inflammatory pain in rats. Methods Sixty female SD rats weighing 150-180 g in which intrathecal (IT) catheters were successfully placed without complication were randomly divided into 5 groups (n= 12 each): group Ⅰ sham operation; group Ⅱ sham operation + IT anti-BDNF antibody; group Ⅲ inflammatory pain; group Ⅳinflammatory pain + IT control IgG and group Ⅴ inflammatory pain + IT anti-BDNF antibody. Inflammatory pain was induced by injecting complete Freund's adjuvant (CFA) into ankle joint cavity of left hindpaw, while in sham operation group equal volume of normal saline was injected instead of CFA. Anti-BDNF antibody or control IgG 15 μg/10 μl was injected IT once a day for 3 days after inflammatory pain was induced. Paw withdrawal latency to thermal stimuli (PWTL) was measured one day before and at 3, 5, 7, 10 and 14 d after inflammatory pain was induced. The rat was sacrificed on 3 rd day of IT anti-BDNF antibody or control IgG injection. The lumbar segment L4-6 of the spinal cord was removed for detection of the expression of BDNF and p-ERK1/2 by immunohistochemistory and Western blot. Results Intra-articular CFA injection significantly increased the expression of BDNF and p-ERK1/2 in the spinal cord in group Ⅲ as compared with sham-operated animals in group Ⅰ . IT antiBDNF antibody injection significantly suppressed the expression of BDNF and p-ERK1/2. PWTL was significantly shortened after intra-articular CFA injection in group Ⅲ as compared with group Ⅰ . IT anti-BDNF antibody reversed the inflammation-induced thermal hyperalgesia in group Ⅴ but IT control IgG did not. Conclusion BDNF in the spinal cord may be involved in inflammatory pain through p-ERK1/2 signal transduction pathway.

Objective To revise looming maladaptive style questionnaire(LMSQ-R) and examine its reliability and validity.Methods 284 undergraduates were measured preparedly with LMSQ-R,281 university students participated in a retest,using LMSQ-R,fear of negative evaluation scale (FNE),Beck anxiety inventory (BAI),Beck depression inventory (BDI).Results ①The item distinguish analysis was acceptable.②Reliability analysis confirmed that Cronbach α coefficient of LMSQ-R was 0.736,Cronbach α coefficient of the two subscales were 0.593 and 0.636.The test-retest reliability of LMSQ-R ranged from 0.564 to 0.700.③Confirmatory factor analysis suggested that the first order six factor-second order two factor model was perfect according to the evaluation criteria.The correlation coefficient between the two subscales was 0.527,the correlation coefficients among the two subscales and the total score ranged from 0.872 to 0.875.The correlation coefficients among the LMSQ-R and FNE,BAI,BDI ranged from 0.872 to 0.875,the results had statistical significance.Conclusion The revised LMSQ-R shows the satisfactory reliability and validity in university students.It can be used as a useful testing tool of LCS in psychological research.

Objective Toexplore the expressions of interleukin-16 (IL-16), interferon-γ (IFN-γ), CXC chemokine receptor 3 (CXCR3), and CRP and their clinical significance in acute exacerbation chronic obstructive pulmonary disease by observing the changes in these factors in patients with AECOPD. Methods 103 patients with AECOPD and 20 healthy controls were collected. According to the 2013 GOLD guideline, all the patients with AECOPD were divided into4 groups(group A of 21 patients, B of 30, C of 27, andD of 25). Results As compared withthe control group, plasma concentrations of IL-16, IFN-γ, CXCR3. and CRP were significantly increased in the patients with AECOPD (P < 0.01), and as the severity of the disease was elevating, these expression levels were significantly increased.While the expression levels of IL-16, IFN-γ, CXCR3, and CRP levels were significantly reduced after treatment, but they were still higherthan those in the control group (P < 0.05). The expression levels of serum IL-16, IFN-γ, CXCR3, and CRP were significantly correlated in patients with AECOPD. Conclusions Expressions of IL-16, IFN-γ and CXCR3 are significantly increased in AECOPD, which is correlated with disease severity and decreased after treatment, suggesting that these three factors may be associated with the occurrence and development of COPD.

AIM:Mitochondrial DNA (mtDNA) copy number variation (CNV), which reflects the oxidant-induced cell damage, has been observed in a wide range of human diseases .However, whether it correlates with heart failure , which is closely related to oxidative stress, has never been elucidated before .We aimed to systematically investigate the association between leukocyte mtDNA CNV and heart failure risk and prognosis .METHODS: A total of 1 700 hospitalized patients with heart failure and 1 700 age-and gender-matched community population were consecutively enrolled in this observational study , as well as 1 638 ( 96.4%) patients were fol-lowed prospectively for a median of 17 months (12~24 months).The relative mtDNA copy number in leukocyte of peripheral blood or cardiac tissue was measured in triplicate by quantitative real-time PCR method .RESULTS:Patients with heart failure possessed much lower relative mtDNA copy number compared with control subjects (P<0.01), especially for the patients with ischemic etiology (P<0.01).Patients with lower mtDNA copy number exhibited 1.7 times higher risk of heart failure ( P<0.01).Long-term follow-up (median 17 months) showed that decreased mtDNA copy number was significant associated with both increased cardiovascular deaths (P<0.01) and cardiovascular rehospitalization (P<0.01).After adjusted for the conventional risk factors and medications , lower mtDNA copy number were still significantly associated with 50% higher cardiovascular mortality (P <0.05).CONCLUSION:
mtDNA copy number depletion is an independent risk factor for heart failure and predicted higher risk of cardiovascular deaths in patients with heart failure .