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.

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.

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.

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.

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 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 study the effect of nerve growth factor(NGF)on bone fracture healing of inflicted T_(10)spinal cord injury(SCI)complicated with tibia fracture in rats.Methods:Totally 120 rats were randomly divided into tihia fracture group (F group,n=40),T_(10)SCI+tibia fracture group(FS group,n=40),and T_(10)SCI+tibia fracture+NGF group(FSN group,n=40).Four weeks later,the fracture sites in the 3 groups were subjected to CT scanning;the maximum transverse diameter of the fracture ends and the gray scales of non-osseous area were measured;the changes of biomechanics property of the fracture ends were determined by three-point bending test;the bone morphometry,bone density,and histomorphology of callus were determined;the expression of OCN was detected by immunohistochemical method;the osteoblast ultrastructure was observed by TEM and the expression ofⅠ,Ⅱtype collagen were examined by Western blotting.Results:The maximum transverse diameter of F group was less than those of FS group(P

Objective To investigate the role of c-Jun N-terminal kinase (JNK) signal transduction pathway on the up-regulation of the expression of acyl-coenzyme A: cholesterol acyltransferasel (ACAT1) induced by Chlamydia pneumoniae (C. pn), and to discuss the mechanism of macrophages-derived foam cell formation induced by C. pn. Methods C. pn 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 four groups to be incubated continually: control group, C. pn infection group, C. pn and SP600125 (a special JNK inhibitor)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 analysis. The expressions of ACAT1 mRNA and protein were determined by reverse transcriptase polymerase chain reaction(RT-PCR) and Western blot, respectively. Results Compared with the control group, the expressions of ACAT1 mRNA and protein were up-regulated in C. pn infection group [(4.16±0.26) vs. (2.17±0.18), (1.20±0.10)vs. (0.61±0.03), both P<0.05], and C. pn-induced foam cell formation was observed. The expressions of ACAT1 mRNA and protein and the foam cell formation were inhibited by SP600125 in a concentration-dependent manner (r = - 0.92, P<0.05; r= - 0. 96, P<0.05, respectively). Conclusions The up-regulation of ACAT1 expression is induced by C. pn via JNK signal transduction pathway, which is involved in the mechanism of C. pn-induced macrophage-derived foam cell formation.

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 expression and prognostic significance of CD19 in patients with Acute myelogenous leukemia with AML1-ETO positive. Methods: Clinical data of 66 patients AML with AML1-ETO positive who were newly diagnosed from Jan 2010 to Dec 2015 were collected. To retrospectively analyze the relationship between clinical characteristics and expression of CD19,so dose the prognosis. Results:The positive rate of CD19 expressing in AML with AML1-ETO positive was 50. 0%. There were no statistically significant differences in terms of age,gender,hemoglobin,platelet,percentage of bone marrow blasts,accompanied with chromosome ,gene mutations between patients with and without CD19 expression(P>0. 05). The white blood cell count(WBC) of the CD19 negative group was higher than CD19 positive group,while showed significant difference(P=0. 027). Although the relapse-free survival (RFS) of patients with CD19 expression was higher than those without,no significant difference was calculated (P=0. 105). Patients with CD19 expression had superior overall survival ( OS ) compared to those without CD19 expression ( P = 0. 030 ) . Multivariable analysis for OS identified CD19 positivity as an independent predictor associated with better prognosis. Conclusion: The expression of CD19 in AML with AML1-ETO positive may be an indicator associated with better prognosis.