ObjectiveTo investigate the expression of MicroRNA 155 (miR-155) in esophageal squamous cell carcinoma (ESCC) and analyze its correlation with clinicopathological features of ESCC. MethodsThis study included 54 patients with primary ESCC who underwent radical esophagectomy in Department of Thoracic Surgery, Henan Cancer Hospital of Zhengzhou University between January 2010 and November 2012. There were 47 males and 7 females with median age of 61 years (range, 45 to 82 years). Forty patients were in stage Ⅰ or Ⅱ and 14 patients in stage Ⅲ a+b. Expression of miR-155 was determined by SYBR Green qRT-PCR in ESCC tissue and corresponding adjacent normal mucosa in surgical samples from the 54 patients, and its correlation with clinicopathological features was analyzed. ResultsExpression of miR-155 was significantly lower in ESCC tissue than that in adjacent normal mucosa (Z=-4.258, P=0.000).Expression level of miR-155 was significantly correlated with lymph node metastasis (P=0.040), but not significantly correlated with smoking (P=0.430), drinking (P=0.429), age (P=0.769), gender (P=0.671), depth of invasion (P=0.230), differentiation degree (P=0.896) or pTNM (P=0.407) of ESCC. ConclusionUnder-regulation of miR- 155 expression in ESCC tissue may lead to disorders of inflammation response, immune response and relevant tumor suppressor, and may play a significant role in carcinogenesis and progression of ESCC.
ObjectiveTo investigate the expressions of microRNA-155 (miR-155) in different phenotypes of activated macrophages. MethodsThe THP-1 cells underwent polarized activation into M1, M2 or tumor-associated macrophages (TAMs), and the phenotypes were confirmed by flow cytometry. The miR-155 expression was determined by qRt-PCR in M1 macrophages, M2 macrophages and TAMs. ResultsThe miR-155 expression significantly decreased in the M2 macrophages (1.83±0.337, P=0.000), TAMs (1.60±0.233, P=0.000) compared with the M1 (6.580±0.637). The phenotype of TAMs was similar to M2. There was no statistically significant difference between TAMs and M2 macrophages in the expression of miR-155 (P=0.546). ConclusionDifferent expressions of miR-155 in macrophages M1-type and M2-type may be associated with the differentiation or their cellular functions. The phenotypic characteristics TAMs may transform to macrophages to M2-type. And they may have the same functions.
ObjectiveTo explore the effect of five copies hypoxia-responsive element (5HRE) and carcinoembryonic antigen promoter (CEAp) element, and to explore the inhibition effect of lentiviral vectors targeted Ras association domain family 1 isoform A (RASSF1A) gene on SGC7901 human gastric cancer cells. Methods①Expressions of carcinoembryonic antigen (CEA) mRNA and its protein, and RASSF1A protein in SGC7901, MKN28, and MCF-10A cells were detect by real time-PCR (qRT-PCR), immunocytochemistry, and Western blot, to confirm the experimental and negative control cells.②The recombinant vectors of pGL4.20-5HRE-CEAp-Luc were constructed through molecular cloning technique to transfected SGC7901, MKN28, and MCF-10A cells. Each kind of cell was divided into 2 groups:one of them didn't add CoCl2 (normoxia group), and another group added CoCl2 (hypoxia group). Comparison of the fold of activation was performed.③SGC7901 cells were infected by lentiviral vectors of pLV-5HRE-CEAp-RASSF1A (infection group) and negative virus (negative control group), SGC7901 cells without any treatment as blank control group. Then SGC7901 cells of 3 groups were divided into 2 groups:one of them didn't add CoCl2 (normoxia group), and another group added CoCl2 (hypoxia group). The expression of RASSF1A protein was tested by Western blot, and the growth inhibition rate was confirmed by cell counting kit-8 (CCK-8) assay. Comparisons of expression of RASSF1A protein and growth inhibition rate of each group were performed. Results①Results of qRT-PCR, immunocytochemistry and Western blot showed that, SGC7901 cells showed higher expression of CEA mRNA and positive expression of RASSF1A protein than corresponding index of MKN28 cells and MCF-10A cells (P < 0.05), which were assigned as experimental cells; but MKN28 cells showed lower expression of CEA mRNA and negative expression of RASSF1A protein, which were assigned as negative control cells.②In SGC7901 and MKN28 cells transfected recombinant vectors of pGL4.20-5HRECEAp-Luc, compared with normoxia group in the same kind of cell group, the folds of activation in hypoxia group were higher (P < 0.01), but there was no significant difference between the normoxia group and hypoxia group in MCF-10A cells (P > 0.05). In the condition of with or without CoCl2, compared with SGC7901 cells in the same condition, the folds of activation in MCF-10A and MKN28 cells were both lower (P < 0.05); compared with MKN28 cells, the fold of activation in MCF-10A cells was lower (P < 0.05).③Western blot results showed that, in the condition with and without CoCl2, expressions of RASSF1A protein decreased in SGC7901 cells of blank control group and negative control group; weak expressions of RASSF1A protein was observed in SGC7901 cells of infection group when in condition of without CoCl2, but increased when adding CoCl2. But RASSF1A protein didn't expressed in MKN28 cells of blank control group, negative control group, and infection group, whether adding CoCl2 or not. CCK-8 assay result showed that, in SGC7901 cells, the growth inhibition rate of infection group which added CoCl2 was higher than those of other 5 groups (P < 0.05); in MKN28 cells, the growth inhibition rates of infection group and negative group were all higher than those of blank control group, whether adding CoCl2 or not (P < 0.05), but there was no significant difference among the infection group and negative group, whether adding CoCl2 or not (P > 0.05). ConclusionsA new hypoxia inducible and cea-positive tumor-targeting transcriptional regulatory element of 5HRE-CEAp is established successfully, and lentivirus vector of pLV-5HRE-CEAp-RASSF1A can significant inhibit the growth of SGC7901 cells under hypoxia condition.
Objective To monitor the distribution of blood perfusion during aortic arch aneurysm surgery under double arterial lines with single pump. Methods We retrospectively analyzed the clinical data of 37 patients underwent aortic arch repair or reconstruction between September 2012 and April 2014. There were 9 females and 28 males at mean age of 48.1±10.8 years ranging from 19.0-72.0 years.We took double arterial lines with single pump for cardiopulmonary bypass (CPB) during the operation and we monitored the perfusion tube flow of both the upper and lower body by blood flow detector. Cerebral blood perfusion was measured by transcranial cerebral Doppler and near-infrared spectroscopy cerebral oxygen saturation (rSO2). Results The mean CPB time of all 37 patients was 195.8±40.5 minutes ranging from 136.0-277.0 minutes and the mean duration time of selective antegrade cerebral perfusion (SCAP) was 21.6±5.6 minutes ranging from 5.0-35.0 minutes. During cooling and rewarming phases, the part of blood flow through axillary artery cannulation ranged from 31.5% to 40.8% of the whole body perfusion. The blood flow of SACP was increased to 15.0 ml / (kg·min) in 2 patients with significantly lower rSO2 and middle cerebral artery blood flow during SACP, and they had an uneventful recovery process after surgery. There were another 2 patients recorded abnormal situation of rSO2 without interventions. One patient died and the other one recovered with compications of spinal cord. Conclusions The technique of double arterial lines with single pump is reasonable and effective. The cerebral perfusion monitoring is helpful to detect abnormal perfusion during aortic arch aneurysm surgery.