Objective To look for and confirm the downstream regulated gene of microRNA (miRNA)-148a in pancreatic cancer cell line.Methods The target gene regulated by miRNA-148a was predicted through bio-informatics analysis.The plasmid containing desired gene and with luciferase 3'-untranslated region (3'-UTR) reporter was constructed.Pancreatic cancer cells BXPC-3 were transfected with analogs and inhibitors of miRNA-148a by liposomes.The activity of luciferase was measured to determine whether miRNA-148a directly connected with desired gene.The expression level of miRNA-148a was changed in BXPC-3 cells,and the changes of target gene v-verb-b2 erythroblastic leukemia viral oncogene homolog 3 (awian) (ErbB3) expression were detected by Western blot at protein level.The data were analyzed by one way ANOVA.Results There was a conservative binding site of ErbB3 with miRNA-148a detected by bio-informatics analysis,miRNA-148a directly combined with ErbB3 and the activity of luciferase decreased to (25.00+47.00) ％ of the negative control (F=4.66,P＜ 0.01).After miRNA-148a overexpression,the gray value of ErbB3 expression in BXPC-3 cells decreased to (26.16±4.69)％ of control group (F=6.563,P＜0.05).Conclusion miRNA-148a directly targeted and regulated the expression of ErbB3 in pancreatic cancer cell line BXPC-3.

Objective To analyze the suspicious results of serum HBV DNA by fluorescence quantitative PCR and develop appropriate countermeasures in order to improve the quality of detection of HBV DNA.Methods Blood samples of patients from the First Affiliated Hospital of Wenzhou Medical College from 2008 to 2011 were analyzed for HBV DNA by fluorescence quantitative PCR.1969 cases of suspicious results,judged by the rule of review the results of serum HBV DNA combined with the historical results,PCR amplification curve,HBV serum markers and clinical diagnosis,were analyzed and redetected by using of two different reagents,careHBV PCR Kit and careHBV PCR Kit V2,at the same time.The consistency and inconsistency ratio of the results were evaluated.Both the reasons of inconsistent and the undetected rates of careHBV PCR Kit were analyzed.The two reasons for the inconsistent results included the reagent related factors,e.g,showing no amplification curve caused by the false negative and abnormal low efficiency of amplification curve,and the non reagent related factors such as operating pollution and other sample factors.Results There were 115 154 blood samples were detected for HBV from 2008 to 2011 and 1969 samples (1.71％) with suspicious results were redetected.The consistency and inconsistency results were 1588 (80.65％) and 381 (19.35％),respectively.Every year from 2008 to 2011,the percentage of the inconsistent results caused by the reagent related factors were 18.87％,20.23％,51.33％ and 59.57％ respectively,which showed an increasing trend,and the percentage of inconsistent results caused by the nonreagent related factors were 81.13％,79.77％,48.67％ and 40.43％ respectively,which showed a declining trend year by year.The undetected rates of careHBV PCR Kit were 2.49％,4.08％,10.09％ and 14.47％ respectively,showing an increasing trend.Conclusions The redetection for the specimens with the suspicious results by using of different reagents can avoid the blind detection of HBV DNA and reduce the experimental error.All the clinical samples for quantitative HBV DNA including the mutations of HBV gene can be measured accurately and effectively,which is helpful to hepatitis B patients for antiviral therapy.

Objective To investigate the situation of food contamination by lead,cadmium,mercury,copper and aluminum in Ningxia Region.Methods Ten categories of food samples up to 712 copies including dairy,edible mushrooms,eggs,preserved egg,meat,organs,aquatic products,vegetables,grains and pasta from supermarkets of five administrative cities of Ningxia Autonomous Region were selected in 2010.The contents of lead,cadmium and mercury in food were detected in 646 samples of 9 categories of food,and copper content of the 30 meat samples were detected.Aluminum content in 66 samples of flour food was tested.According to the methods of national food safety standards (GB 5009.12-2010) and national food hygiene standards (GB/T 5009.13-2003,GB/ T 5009.15-2003 and GB/T 5009.17-2003),lead,copper,cadmium and mercury in various foods were tested.Aluminum was determined using the method for determination of spectrophotometry.Results The detection rates of lead,cadmium,mercury in the nine categories of food were 52.8％ (341/646),62.1％ (401/646),51.7％ (334/646);the detection rates of copper in the meat was 90.0％ (27/30);the aluminum in flour food was 84.8％ (56/66),respectively.Compared with the national standards,the eligible rates of lead,cadmium,mercury,copper and aluminum were 97.1％ (627/646),98.2％ (539/549),100.0％ (646/646),100.0％ (30/30) and 75.8％ (50/66),respectively.Food contaminated by lead and cadmium were mainly distributed in preserved egg [lead detection rate 9.5％ (2/21)],organs [lead detection rate 6.7％ (2/30),cadmium detection rate 30.0％ (9/30)],vegetables [lead detection rate 9.9％ (8/90)] and a small amount of food products [lead detection rate 4.7％ (7/150)].The levels of lead in fried flour food and fine dried noodles were higher [40.9％ (9/22),36.4％ (4/11)].Conclusions Food contamination by lead,cadmium,mercury and copper is not serious in Ningxia.The monitoring results of lead,cadmium,mercury and copper in food are satisfying in Ningxia.But there is aluminum contamination in flour food.It is important to do the risk evaluation of aluminum contamination in food as quickly as possible.

Objective To evaluate the application of sentinel lymph node biopsy in surgical operation for clinical stage N0 differentiated thyroid carcinoma.Methods One hundred and six patients who were diagnosed clinical stage N0 differentiated thyroid carcinoma in Department of Mammary Gland Thyroid Surgery of Qingdao Women and Children's Hospital between Jan.2012 and Jan.2013 were reviewed retrospectively.Carbon nanoparticles were used to trace sentinel lymph node in the surgery,the patients were divided into the dissection group (46 cases included 2 cases whose sentinel lymph nodes were not detected) and the non dissection group (60 cases) based on the rapid pathological results.The operation index,complications incidence and short-term outcomes were compared.Result The operative time,intraoperative blood loss and the faulty excision of parathyroid gland in dissection group is much higher than those in non dissection group (P ＜0.05).The incidence of provisionality hypoparathyroidism in dissection group is much higher than those in non dissection group (P ＜ 0.05).There were no permanent hypoparathyroidism and damage of laryngeal nerve in the two groups.Postoperative outpatient follow up was 30 months,no significant differences were seen in local regional recurrence rates(6.5％ vs 10％) between the two groups (P ＞ 0.05).There were no distant metastasis occurred in the two groups.Conclusions Lymph node metastasis can be found by sentinel lymph node biopsy,it is helpful for us to decide whether dissect central lymph node or not.Carbon nanoparticles is safe,which is helpful to protect the parathyroid gland.

Objective To investigate the effect of adenosine A2A receptor on pituitary-adrenal axis response in acute phase of moderate craniocerebral trauma.Methods Eighteen adenosine A2A receptor knock-out mice in a C57BL/6 background and another eighteen their wild-type littermates were divided into normal control group and craniocerebral trauma for 4 hours group,and craniocerebral trauma for 24 hours group according to random number table,with siμ mice per group.Plasma levels of adrenocorticotropic-hormone (ACTH) and corticosterone at hours 4 and 24 postinjury were determined using ELISA method.Results At 4 and 24 hours,brain water content in wild-type mice [(80.950 ± 0.184) ％,(82.178 ± 0.255)％ respectively] was higher than that in gene knock-out mice [(80.006 ± 0.199)％,(81.091 ± 0.295)％ respectively,P ＜ 0.01].Besides,brain water content in both wild-type and gene knock-out mice increased after injury (P ＜ 0.01).Plasma levels of ACTH and corticosterone were higher in geneknock-out sham mice than in wild-type sham mice [(120.214 ± 2.472) ng/L vs (91.767 ±7.395) ng/L,(27.814 ±0.888) μg/L vs (11.430 ±0.644) μg/L respectively,P ＜0.0l].At 4 and 24 hours,plasma levels of ACTH [(174.776-± 5.040) ng/L,(189.613 ± 4.802) ng/L respectively] in geneknock-out mice showed a higher increase than those in wild-type mice [(119.594 ± 6.945) ng/L,(124.93-± 11.001 7) ng/L respectively,P ＜ 0.05].Moreover,plasma levels of corticosterone [(40.138 ±-0.805) μg/L] at 4 hours and [(37.440-0.485)μg/L] at 24 hours in gene knock-out mice showed a same result as compared with that in wild-type mice [(19.702 ± 0.804) μg/L,(17.602 ± 0.743) μg/L respectively,P ＜ 0.05].Conclusions Knock-out of adenosine A2A receptor increases the release of ACTH and corticosterone in acute stage of moderate craniocerebral trauma and promotes pituitary-adrenal stress response.This may provide a novel explanation for the neuroprotective effect of A2A receptor deficiency.

Objective To investigate the circulating microRNAs carrier in pancreatic cancer.Methods Pancreatic cancer cell lines SW1990 and BxPC3 were routinely cultured and serum of 6 patients with pancreatic cancer and 6 healthy subjects (control group) were collected.Serum and pancreatic cancer cell line supernatant microvesicles (MV) were obtained by gradient centrifugation.The expression of Ago2,CD63 was detected by Western blotting,the expression of microRNA in microvesicle section and microvesicle-free section in serum was detected by using quantitative PCR method.Results The supernatant MV of pancreatic cancer cell lines expressed Ago2,CD63 protein,and these MV carried different microRNAs in different cell lines.In the serum of pancreatic cancer and control group,miR-20a,miR-21,miR-24,miR-25,miR-191,miR-483-5p were detected,but the quantity was relatively higher in MV section,and the expression of microRNAs in pancreatic cancer's MV was inconsistent with that of control group.The expression of miR-20a,miR-24,miR-191 in pancreatic cancer group was (2.93 ± 0.29),(2.73 ± 0.46),(2.39 ± 0.51) times as much as those in control group,and the difference between the two groups was statistically significant (F =75.97,25.80,12.94,P ＜ 0.05 or ＜ 0.01).Conclusions The main circulating microRNAs carrier in pancreatic cancer is microvesicle.

Objective To compare the protective effect of double dose trimetazidine versus standard dose in patients with unstable angina and received PCI. Methods From September 2014 to June 2015,150 unstable angina pectoris patients who underwent PCI in our hospital were enrolled in this study. All the patients were randomized into two groups:the study group (patients given trimetazidine 40 mg tid 24 hours before operation) and the control group (patients given trimetazidine 20 mg tid 24 hours before operation). All patients received standard secondary prevention therapies of coronary heart disease. Comparison in the rates of angina pectoris attacks, ECG changes after PCI, levels of cardiac troponin I ( cTnI), myoglobin (Mb), creatine kinase isoenzyme ( CK-MB) and hs-CRP before and 12 hours after PCI was investigated between the 2 groups. Results Baseline data were comparable in the 2 groups. Rates of adverse drug reaction were lower in the control group, but the difference was not statistically significant (6. 7% vs. 2. 7% , P = 0. 439). There was no statistical difference between the two groups before PCI in terms of levels of cTNI and hs-CRP but was significantly lowered in the study group after PCI as compared to the control (both P < 0. 05 after PCI). The levels of CK-MB and Mb in the study group were lower than those in the control group after PCI, but the difference was not statistically significant. The rates of angina pectoris attacks and the changes in ST-T were less in the study group as compared to the control group without statistical significance. Conclusions Unstable angina pectoris patients receiving double dose of trimetazidine before PCI may enhance myocardial protection and reduce PCI related myocardial injury.

ObjectiveTo investigate the mechanism of dexamethasone (Dex) in inhibiting monocyte adhesion and phagocytose function.Methods Under the stimulation of phorbo1-12-myristate-13-acetate (PMA),U937 monocytes cultured in vitro were treated with Dex and Fasudil respectively.The adhesion rate of U937 monocles to human umbilical vein endothelial cells (HUVECs) and their phagocytic ability of India ink were studied.The protein content and activity of rho-associated coiled-coil protein kinase 1 ( ROCK1 ) as well as the effects of mifepristone and cycloheximide on Dex were determined.ResultsBoth DEX and Fasudil could significantly inhibit the adhesion tate and phagocytosis of U937 cells stimulated by PMA and suppressed the activity of ROCK1.While mifepristone and cycloheximide could not alter these effects of DEX.ConclusionDEX interferes with the adhesion and phagocytosis function of U937 cells by inhibiting ROCKI activity.

Objective:To investigate the feasibility of detecting excision repair cross-complementing 1(ERCC1)and ERCC2 in peripheral venous blood instead of cancer tissues from esophageal squamous cell carcinoma patients. Methods:The expressions of ERCC1 and ERCC2 mRNA were detected by using RT-PCR in 39 cases of peripheral venous blood samples, esophageal squamous cell carcinoma tissues, and adjacent normal tissues. ELISA was used to determine the levels of ERCC1 and ERCC2 proteins in serum. The periphe-ral blood from 10 healthy volunteers was used as control. Results:Expression levels of ERCC1 and ERCC2 mRNA and protein were significantly higher in peripheral blood from healthy control than those in esophageal carcinoma patients (P<0.05). There was a positive correlation between the expression of ERCC1 and ERCC2 mRNA in peripheral blood and cancer tissues (P<0.01). Conclusion:The expression levels of ERCC1 and ERCC2 mRNA in peripheral blood can indirectly reflect their expression levels in human esophageal squamous cell carcinoma tissues.

Objective To analyze the characteristics of preoperative CTO lesions by coronary CT angiography (CCTA) and to compare the lesion characteristics and clinical data of patients with subsequent vs failed PCI.Methods A total of 113 patients were randomly selected and 116 vessels were analyzed by CCTA before PCI.The patients were further investigated as PCI success group vs PCI failure group according to their PCI result.Multivariate logistic regression analysis was used to determine the factors that affected the success of CTO intervention.The ROC curve was used to determine and evaluate the CT-CTO score and J-CTO score for diagnostic efficacy.Results The success rate of PCI was 55.2％.64 lesions were successfully opened,with the success rate of 72.4％.The prevalence of smoking in patients in the PCI failure group was significantly higher than that in PCI success group (65.4％ vs.42.2％,P ＜ 0.05).There were no significant differences between the two groups in age,gender,history of hypertension,diabetes mellitus,and myocardial infarction(P ＞ 0.05).Statistical differences were observed between the PCI success group and the PCI failure group in the presence of occlusion segment head-end bifurcation,occlusion severe incision,severe calcification (calcification ≥ 180°),occlusion segment length ≥ 20 mm,occlusion of calcification lesions,occlusion segment distal shape of the unambiguous of fiber cap shape of the distal occlusion segment under CCTA(P ＜ 0.05).In the PCI failure group,approximately 17.3％ of the patients had previous attempt to open the CTO lesions,which were higher than the PCI success group (9.4％).However,The difference was not statistically significant (P ＞ 0.05).Multivariate logistic regression analysis showed that the unambiguous distal fibrous cap of the occlusion segment and the occlusion of the proximal branch and the occlusion length ≥20 mm were the main factors affecting the failure of CTO intervention.In terms of prediction,the predictive value 30 CT-CTO score yielded a higher area under the ROC curve than that of the J-CTO score (0.8776 vs 0.7387,P ≤ 0.05).Conclusion CT angiography can predict the success rate of intervention for CTO lesions.Compared with J-CTO score,CT-CTO score has a higher predictive value.Unambiguous fiber cap shape,occlusion segment head end bifurcation,occlusion segment length ≥20 mm were the independent risk factors that affecting the success of CTO operation.