Objective To construct plvx-cyclooxygenase-2(COX-2)-DsRed and establish breast cancer cell line MCF7 which overexpressed COX-2, to explore the effect of COX-2 on breast cancer cell.Methods The full-length COX-2 PCR product was obtained by total COX-2 PCR primers and COX-2 cDNA vector.After the PCR product and lentiviral vector plvx-DsRed-Monomer-N1 were cut simultaneously by restriction enzyme BamH1 and Xholl, they were connected and sequenced, to get lentiviral vector plvx-COX-2-DsRed.After selected by puromycin, overexpressed COX-2 breast cancer cell line MCF7-plvx-COX-2-DsRed was obtained.The stable cell line was verified by real time PCR and Western blot.The differences of proliferation ability between stable cell line and normal one were compared by colony formation test and Western blot.Results The lentiviral vector plvx-COX-2-DsRed and stable cell line MCF7-plvx-COX-2-DsRed after selecting were obtained.COX-2 expression level of the stable cell line was 75.29 times as high as that of MCF7, and 64.91 times as high as that of cell line MCF7-plvx-DsRed-Monomer-N1 by PCR assay (P ＜ 0.05), which was consistent with the results of Western blot and microscope photo.MTT results showed that cell line MCF7-plvx-COX-2-DsRed had grown faster than cell line MCF7 and MCF7-plvx-DsRed-Monomer-N1 from the 2nd day (P ＜ 0.05), which was accordant with colony formation assay.MCF7-plvx-COX-2-DsRed cell line had higher c-myc expression and lower β-catenin expression than MCF7 cell and cell line MCF7-plvx-DsRed-Monomer-N1 detected by Western blot relative quantification (P ＜ 0.05).Conclusion The plvx-COX-2-DsRed lentiviral vector and cell line MCF7-plvx-COX-2-DsRed are successfully constructed.COX-2 can increase proliferation of MCF7 cells through up-regulating the expression of c-myc.

Objective To further understand the diagnosis,clinical features and prognosis of myeloid neoplasms with erythroblast more than 50％ of bone marrow(BM) nucleated cells in the WHO Classification(2016) by analyzing the clinical data,diagnosis and prognosis of 3 patients with myeloid leukemia.Methods The ages,medical histories,symptoms and signs,and laboratory examinations from 3 patients with myeloid neoplasms whose erythroblast cells were more than 50％ of BM nucleated cells when newly diagnosed were collected.Then,they were diagnosed with the WHO Classification criteria(2008) and the WHO Classification criteria(2016),respectively,and their prognosis was evaluated with the revised International Prognostic Scoring System(IPSS-R).Results According to the WHO Classification criteria(2008),all of 3 patients were diagnosed as acute erythroid leukemia(AEL).However,according to the WHO Classification criteria(2016),2 patients were diagnosed as myelodysplastic syndrome with excess blasts-2(MDS-EB-2),and 1 was diagnosed as acute myeloid leukemia(AML) with maturation.Moreover,their prognostic scores were also different.The former two patients were older men with significant dysplasia and complex genetic abnormalities,and had poor prognosis,while the latter was a middle-aged woman with no obvious dysplasia and genetic abnormalities,and had medium prognosis.Conclusion The WHO Classification(2016) is more reasonable than the WHO Classification(2008),which tends to focus more on the different biological characteristics of diseases,and may better distinguish two types of diseases with different clinical features and prognosis.

Objective By analyzing the perioperative management in our hospital to explore the clinical effect and safety of single kidney transplantation from deceased juveniles' donors.Methods We retrospectively analyze 86 cases of kidney transplantations from deceased juveniles' donors in our hospital from 2007 December to 2015 August.Results The success rate of the operations was 100％.The postoperative complications occurred as fellows:7 cases of acute rejection (8.14％);10 cases of drug intoxication (11.62％);21 cases of DGF (24.44％),4 cases of leakage of urine (4.65％),7 cases of lung infection (8.14％).Two cases (2.32％) died after the operation because of serious lung infection,and by corresponding treatment 47 cases recovered after 2-4 weeks.The creatinine level in 37 cases without any complications was 131.88 ± 44.20 μmol/L during discharge.Conclusion With strict selection,the organ from a deceased juvenile donor is safe and practicable.

【Objective】 To investigate the effect of prenatal stress (PS) on the profile changes of cardiac metabolites in offspring rats and to analyze the potential role of key differentiators in key differential signaling pathways. 【Methods】 UHPLC-Q-TOF/MS analysis was used to detect the changes of metabolite profile in the heart tissues of offspring rats in control group and PS group. KEGG pathway annotation and analysis were used to screen out metabolic differences in key signaling pathways and quantify their expression levels so as to predict the potential function of these key molecules in the effect of PS on the heart tissues of offspring rats. 【Results】 Compared with the control group rats, the signaling pathways in the PS offspring rats’ heart tissue that changed significantly included biosynthesis of amino acids, purine metabolism, pyrimidine metabolism, alanine, aspartate and glutamate metabolism, cAMP signaling pathways, arginine biosynthesis, GABAergic synapses, glutamate synapse, nicotine addiction, and regulation of actin cytoskeleton. Among them, the levels of L-glutamine, pseuduracil, uric acid, xanthine, 2’-deoxyadenosine 5’-monophosphate, cytosine 3’-monophosphate, and cytosine 5’-monophosphate were upregulated, while the level of argininosuccinic acid was downregulated, which enriched in purine metabolism, pyrimidine metabolism, and arginine biosynthesis pathway. 【Conclusion】 PS leads to abnormal changes of L-glutamine, pseuduracil, uric acid, and xanthine in the heart tissue of offspring rats, and PS may be a high risk factor for cardiovascular diseases in offspring rats.

Purpose To assess the feasibility of nuclear score combined with ultrasound features on indeterminate thyroid nodules in fine needle aspiration cytological diagnosis.Methods A consecutive cohort of 119 thyroid fine-needle aspiration(FNA)specimens with indeterminate diagnosis(TBSRTC cate-gory Ⅲ-Ⅴ)and available histopathologic follow-up was evalua-ted.Original sonographic images and cytological slides of each nodule were reviewed by two experienced sonographers and two experienced cytopathologists respectively,and evaluated jointly to obtain consensus.The identification of the optimal cut-off points of simplified nuclear score and ultrasound features score for the diagnosis of malignancy or low-risk neoplasm were evalu-ated using the receiver operating characteristic(ROC)curves and the assessment of the area under the ROC curve(AUC).The specificity,sensitivity,positive predictive value(PPV)and negative predictive value(NPV)of nuclear score and US score were evaluated from crosstabs based on Cut-off points and signif-icance were calculated.The diagnostic accuracy of simplified nuclear score combined with ultrasound features was estimated by Logistic regression and ROC curve analysis.Results Nucle-ar grooves,intra-nuclear inclusions and chromatin clearing were more common in malignancy/low-risk neoplasms(P = 0.001,0.013 and 0.001 respectively).A Cut-off point of≥2 for the simplified nuclear score was sensitive for defining malignancy/low-risk neoplasm,and the sensitivity was 99.1%.Nodule height greater than width,absent halo sign,irregular margin and absent intranodular vascularization were more common in malig-nancy/low-risk neoplasms(P = 0.003,0.007,0.003 and 0.004 respectively).When the Cut-off point was set as≥5,the PPV and specificity for defining malignancy/low-risk neoplasm were 95.9%and 78.6%respectively.ROC curve analysis showed that the diagnostic accuracy of nuclear score combined with US score(0.873,95%CI = 0.777-0.970)was higher than that of nuclear score alone(0.817,95%CI = 0.687-0.948,P =0.092)or US score alone(0.758,95%CI 0.637-0.879,P =0.047).But no significant difference was detec-ted between the combined score and the nuclear score.Conclu-sion The addition of simplified nuclear score and US score to FNA cytology can increase the diagnostic accuracy of cytology in indeterminate categories.