AIM:To study the effect of acyl coenzyme A:cholesteryl acyltransferase 1(ACAT1)antisense oligonucleotides on the formation of foam cells(FC).METHODS:THP-1 cells were cultured and differentiated into macrophages(MP)by phorbol myristate acetate(PMA).Over-expressing ACAT1 gene THP-1 cells were constructed.The ACAT1 antisense and missense oligonucleotides conducted by LipofectamineTM 2000 were incubated with above cells.Ac-LDL was added 6 h later and incubated for 24 h.The expression of ACAT1 protein was detected by Western blotting.The ACAT activity was measured by quantifying the incorporation of 1-14C oleoyl CoA into cholesteryl esters.The formation of foam cells was detected by oil red O staining.RESULTS:The ACAT1 antisense oligonucleotides inhibited the activity of ACAT in macrophages and over-expressing ACAT1 gene THP-1 cells.It also inhibited the formation of foam cell in macrophages and over-expressing ACAT1 gene THP-1 cells with lipid loading.The missense oligonucleotides did not show the inhibitory effects.CONCLUSION:The ACAT1 antisense oligonucleotides inhibit the activity of ACAT and the formation of foam cells.

Objective To observe the expression of MTA1 and ER in breast cancer,the correlation of the two factors and with the invasive capability of breast cancer.Methods The expression of MTA1 and ER in normal breast,precancerous lesions and breast cancer tissues was detected by using nucleic acid hybridization in situ(ISH) and immunocytochemistry(IHC) methods,and their correlation was analyzed by Spearman method.ResultsThe expression of MTA1 and ER was higher in ISH than in IHC.The mRNA expression of MTA1 in normal breast tissue,precancerous lesions and breast cancer tissne was 12.2%,33.3%,and 81.1% respectively,and the expression by IHC was 11.1%,31.1% and 72.2% respectively.The mRNA expression of ER in normal breast tissue,precancerous lesions and breast cancer tissue was 83.3%,61.1% and 37.8%,respectively,and the expression by IHC was70.9%,56.7% and 35.6% respectively.The positive expression of MTA1 was higher in ER-negative patients than that in ER-positive ones(86.2﹪vs.46.9﹪).ConclusionsCombined ISH and IHC detection can improve the detection rate of MTA1 and ER.With advancement of the disease and lowering of tumor differentiation,the expression of MTA1 gradually increases,while expression of ER decreases and even disappears.The expression of MTA1 is negative in relation to that of ER(the coefficient is-0.466).MTA1and ER could be important molecular markers for the prognosis and therapy of breast cancer.

Objective To investigate the application value of BIS monitoring and Ramsay score in the prevention of unplanned tracheal extubation in ICU patients.Methods 93 patients were enrolled in this study,they were divided into the experimental group(47 cases)and the control group(46 cases) using random number method.They received sedation regimens with BIS monitoring and Ramsay score or Ramsay score respectively.Occurrence rate of unplanned extubation was compared between the two groups.Results The occurrence rate of unplanned extubation was significantly lower in the experimental group than that of the control group.Conclusions BIS monitoring and Ramsay score is a suitable ways for the management of sedation of intubated patients.

Objective To investigate the effect of Cimetidine on the adhesion between colorectal cancer LOVO cells and endothelial ECV304 cells;and study whether Cimetidine can inhibit the expression of E-selectin in ECV304 cells. Methods Cellular uptake of rose Bengal stain was used to measure the adhesion between LOVO cells and ECV304 cells. Enzyme-linked immunosorbent assay(ELISA)and flow cytometry (FCM, indirect fluorescence staining and labeling)were used to detect the expression of E-selectin. Results ECV304 cells were exposed to different concentrations of Cimetidine. Both the adhesion between LOVO cells and ECV304 cells and the levels of E-selectin significantly decreased with increasing concentration of Cimetidine(P =0. 001 and 0. 001 respectively). Conclusion Cimetidine can inhibit the adhesion of human colorectal cancer LOVO cells on endothelial ECV304 cells by blocking E-selectin expression. Our observations indicated a potential of anti-adhesion therapy using Cimetidine in cancer treatment.

In order to explore the effect and mechanisms of tumor necrosis factor-alpha (TNF-alpha) on the activity of the acyl coenzyme A: cholesteryl acyltransferase (ACAT), THP-1 monocytes were cultured and induced to differentiate into macrophages with phorbol ester. TNF-alpha (60 ng/mL) was added at different time points into the macrophage-containing medium and the ACAT enzyme activity was measured by quantifying the incorporation of [1-(14)C] oleoyl CoA into cholesteryl esters. The expression of ACAT-1 protein and mRNA was respectively detected by Western blotting and RT-PCR in THP-1 macrophages 24 h after treatment with TNF-alpha (60 ng/mL). The results indicated that ACAT activity in THP-1 macrophages treated with TNF-alpha was increased in a time-dependent manner. The expression levels of ACAT-1 protein and mRNA were significantly increased in THP-1 macrophages after treatment with TNF-alpha (P<0.05). It was suggested that TNF-alpha could increase the activity of ACAT in THP-1 macrophages by up-regulating the expression of ACAT-1 gene.

AIM: To investigate the signal transduction mechanism of Chlamydia pneumoniae (Cpn) in down-regulating the expression of ATP binding cassette A1 (ABCA1) and ATP binding cassette G1 (ABCG1),the key molecules in cholesterol efflux and atherogenesis,from THP-1-derived macrophages. METHODS: Cpn was propagated in Hep-2 cells. THP-1 monocytes were induced into macrophages by 160 nmol/L phorbol myristate acetate (PMA) for 48 h,and were randomly allocated into 4 groups to incubate continually: control group,50 mg/L low density lipoprotein (LDL); Cpn infection group,Cpn (1×10~6 IFU) and 50 mg/L LDL; Cpn and SP600125 (a special JNK inhibiter) group,THP-1 macrophages were previously treated with different concentrations (1-20 μmol/L) of SP600125 for 1 h,and then infected with Cpn (1×10~6 IFU) and 50 mg/L LDL; SP600125 group,SP600125(20 μmol/L)and 50 mg/L LDL. The expressions of ABCA1/ABCG1 and peroxisome proliferator-activated receptor γ (PPARγ) from each group were detected then. The cholesterol efflux was detected by enzyme-fluorescence. The expressions of ABCA1/ABCG1 and PPARγ mRNA and protein were determined by RT-PCR and Western blotting,respectively. RESULTS: Cpn not only down-regulated the ABCA1/ABCG1 expression,but also down-regulated the expression of PPARγ,which regulated the ABCA1/ABCG1 genes transcriptions. However,the mentioned effects of Cpn infection were restrained by the special JNK inhibitor SP600125 in a dose-dependent manner. CONCLUSION: Chlamydia pneumoniae may down-regulate ABCA1/ABCG1 expression from THP-1-derived macrophages via JNK-PPARγ signal transduction pathway.

Objective:To investigate the effects of Chlamydia pneumoniae(Cpn) on SR-A1 and CD36 expression in THP-1-derived macrophages and role of c-Jun NH_2-terminal signal transduction pathway in the process.Methods:Cpn was propagated in Hep-2 cells.THP-1 monocytes were induced into macrophages by 160 nmol/L phorbol myristate acetate(PMA)for 48h,and were randomly allocated into four groups to be incubated continually: control group;Cpn infection group;Cpn and SP600125(a JNK inhibiter)group and SP600125 group.Lipid droplets in cytoplasm were observed by oil red O staining.The contents of intracellular cholesterol ester were detected by enzyme-fluorescence.The expression of SR-A1 and CD36mRNA and protein were determined by RT-PCR and Western blot, respectively. Results:THP-1-derived macrophages infected with Cpn resulted in large accumulation of lipid droplets and foam cell formation when co-cultured with LDL.Meanwhile,the expression of SR-A1 mRNA and protein were up-regulated by Cpn infection (P<0.05).However,the expressions of CD36 mRNA and protein in THP-1-derived macrophages infected with Cpn were unchanged.Moreover,the up-regulation of SR-A1 and foam cell formation induced by Cpn could be restrained by the JNK inhibiter SP600125 in a dose-dependent manner,and SP600125 had little impact on the expression of CD36 in THP-1-derived macrophages infected with Cpn.Conclusion:The up-regulation of SR-A1 but not CD36 expression is involved in mechanisms of Cpn inducing foam cell formation.And Chlamydia pneumoniae up-regulates the expression of SR-A1 via the JNK signal transduction pathway.This may be a novel mechanism for the foam cell formation induced by Cpn.

Objective To evaluate the influence of gestational diabetes mellitus (GDM) on maternal and perinatal outcomes in twin pregnancies. Methods We retrospectively analyzed the clinical features of both twin and singleton pregnancies, which delivered in Guangzhou Women and Children's Medical Center between January 1, 2012 and December 31, 2013. The twin pregnancies were divided into two groups:those with (GDM-T, n=51) and without GDM (non-GDM-T, n=130), which were matched by maternal age and delivery time (within one month) in a ratio of 1∶2 among singleton pregnancies with (GDM-S, n=102) and without GDM (non-GDM-S, n=102), respectively. The differences of adverse maternal and perinatal outcomes among these four groups were examined. The overall assessment of pregnancy outcomes was completed using Delphi method. Statistical analysis was performed with one-way analysis of variance, t test, Kruskal-Wallis test, rank test, Chi-square test or Fisher's exact test. Results (1) When compared to GDM-S and non-GDM-S group respectively, less women conceived with the help of assisted reproductive technology, higher proportion of women underwent and gestational age at delivery tend to be earlier in GDM-T and non-GDM-T group (all P<0.01). In oral glucose tolerance test,the fasting blood glucose level of GDM-T group was higher than the other three groups (F=21.716, P<0.01), the glucose levels at 1 and 2 h were higher than non-GDM-T and non-GDM-s respectively (both P<0.01), but no significant difference was found when compared with GDM-S group (P>0.01). Similarly, no significant difference was found in prenatal glycosylated hemoglobin value between GDM-T and GDM-S group (P>0.01). (2) There was no significant difference in the incidences of hypertensive disorders of pregnancy, anemia, premature rupture of membranes, oligohydramnios, placental abruption, postpartum hemorrhage, asphyxia neonatorum, small for gestational age, hypoglycemia of newborn, hyperbilirubinemia of newborn and perinatal death between GDM-T group and the other three groups(all P>0.01). Higher incidences of hypertensive disorders of pregnancy and postpartum hemorrhage were shown in the GDM-T group than in the GDM-S and non-GDM-S groups, respectively (both P<0.01). The incidences of preterm birth in GDM-T and non-GDM-T group were both higher than that in GDM-S and non-GDM-S, respectively [54.9%(66/102), 53.8%(140/260), 5.0%(10/102) and 3.0%(6/102), all P<0.01], while no significant difference was found between GDM-T and non-GDM-T group (P>0.01). (3) The overall assessment of pregnancy outcomes did not show any difference between GDM-T group and the other three groups (χ2=6.707, P>0.01). However, the score for fetal outcomes in the GDM-T group was higher than in the GDM-S and non-GDM-S group, but lower than in non-GDM-T group [M(Q)=1.0(2.3), 0.0(3.0), 0.0(0.0), 1.0(2.8) score, χ2=122.818, P<0.01]. Conclusions GDM does not increase the risk of adverse pregnant outcomes in twin pregnancies.

Objective To investigate the mechanisms of Chlamydia pneumoniae (C. pn)-induced foam cell formation, the expression of ATP binding cassette transporter AI ( ABCA1 ) and perexisome prolif-erator-activated receptor γ (PPARγ) were examined. Methods THP-1 monneytes were induced into mac-rophages after the addition of 160 nmol/L phorbol myristate acetate (PMA) for 72 h. THP-1-dorived macro-phages when co-cuhured 50 mg/L low density lipoprotein (LDL) were designated randomly in four groups: control (uninfected) group, C. pn infection group, rosiglitazone + C. pn infection group and rosiglitazone group. Lipid droplets in cytoplasm were observed by oil red O staining. The contents of intracellnlur choles-terol ester were detected by enzyme-flnoreseence. The expression of ABCA1, PPARγ, mRNA and protein were determined by RT-PCR and Western blot, respectively. Results C. pn down-regulated the expression of ABCA1, PPARγ at mRNA and protein levels in a concentration-dependent manner in THP-1-derived mac-rophages when co-incubated with LDL. Resiglitazone not only concentration-dependently alleviated the down-regulation of ABCA1 expression by C. pn infection (P<0.05), but also markedly suppressed the accumula- tion of lipid droplets and cholesteryl ester by C. pn at higher concentrations ( 10 and 20 μaol/L). Condu-sion C. pn induces foam cell formation by down-regulating the expression of ABCA1 via PPART pathway, which may provide a new evidence for the development and progression of atherosclerosis initiated by C. pn infection.

Objective To explore the more effective measures for the prevention of deep vein thrombosis (DVT ) by comparing the different efficacy between the multimodal strategy and liberal measures .Methods From July 2011 to June 2013 ,medical records of 289 patients who had accepted total knee replacement (TKR) were collected .Patients were divided into two groups according to whether exploring to the multimodal strategy or liberal measures after TKR .The multimodal strategy consisted of sequentially used perioperative intermittent pneumatic compression ,intermittent pneumatic compression ,and postoperative continues femoral nerve block analgesia .Patients in group A were treated during July 2011 to June 2012 ,and accepted liberal measures for the prevention of DVT .Patients in group B were treated after June 2012 ,and accepted multimodal strategy .The data of each group were collected for statistical analysis on the following aspects :DVT occurrence rate ,DVT distribution ,age ,gender ,body mass index ,disease ,operation duration ,volume of blood loss and transfusion during operation phase ,drainage volume after TKR .Results The DVT occurrence rate of patients in group B were significantly lower than that of patients in group A (P<0 .05) .No significant difference were foun-ded in the DVT distribution between the two groups(P>0 .05) .Conclusion The multimodal strategy consisted of sequentially used perioperative intermittent pneumatic compression ,postoperative low molecular weight heparin ,and postoperative continues femoral nerve block analgesia is more effective than liberal measures for the prevention of DVT .