In this paper, we designed and evaluated a duplex detection strategy for microRNAs (miRNAs) using universal probe-based target-triggered double hybridization and fluorescent microsphere-based assay system (xMAP array). In the absence of target miRNA, reporter DNA cannot hybridize stably with the immobilized capture DNA due to its low melting temperature. Only after adding target miRNA, can reporter probe hybridize with capture probe to form a stable three-component complex. This targettriggered stable hybridization makes this method possible for highly selective and sensitive detection of multiple miRNAs. We exemplified a quantitative detection of duplex miRNAs with a limit of detection of 40 pM. The xMAP array platform holds the potential of extending this approach to simultaneous detection of up to 100 miRNA targets. Considering the simplicity, rapidity and multiplexing, this work promised a potential detection of multiple miRNA biomarkers for early disease diagnosis and prognosis.

Objective:? N oxalo L ?,? diaminopropionic acid were isolated from the powder of red ginseng. its physiological characteristics were studied. Method:This material is obtained by CM Sephadex C 25 and FPLC MonoQ. We have examined the effect of ? N oxalo L ?,? diaminopropionic acid on contraction of artery of guinea pig. Result:When histamin and adrenalin do not exist, there is neither contraction nor relaxation. It can strengthen induction of contraction of artery of guinea pig, but it has no such effect on adrenalin. Besides, it doesn't induce plaque coagulation through adrenalin, ADP blood coagulation enzyme and collagen. It doesn't suppress the transformation from tension peptide of blood vessel to enzyme.Conclusion:Hemostatic effect of ? N oxalo L ?,? dlaminopropinomie acid may caused by vesseles contraction through promoting Histamine.

Objective To investigate the effect of comprehensive nursing intervention on the quality of life of patients suffering from hypertension. Methods A total of 120 cases of essential hypertension were evenly randomized into observation group and control group. Both groups received conventional antihypertensive drug and nursing, and additionally, the observation group was given intervention of comprehensive nursing including dietary regulation, exercise training, emotional care and medication instruction. The intervention lasted 3 months. Before and after the intervention, traditional Chinese medicine ( TCM) symptom classifying list and World Health Organization ( WHO) QOL-BRIEF were applied to evaluate the therapeutic effect of the two groups. The changes of blood pressure, clinical manifestation and the quality of life were also compared between the two groups. Results (1) After intervention, the systolic pressure and diastolic pressure were both improved in the two groups ( P<0.05 or P<0.01 compared with those before treatment) , and the improvement in the observation group was superior to that in the control group (P<0.01). (2) After intervention, the scores of TCM symptom scores and QOL-BRIEF total scores were improved in the two groups ( P<0.05 or P<0.01 compared with those before treatment) , and the improvement in the observation group was superior to that in the control group ( P<0.01). Conclusion TCM comprehensive nursing intervention can effectively relieve hypertension and improve the quality of life of hypertension patients.

Hepatocellular carcinoma (HCC) is the most common malignancy in the liver, for which surgical operation remains the primary treatment. However, the surgical treatment is associated with low resection rate and high recurrence rate, which drive studies on the molecule mechanism of initiation, metastasis, and invasion of HCC, in order to develop more effective early diagnosis and treatment methods. By reviewing related literature, this article summarizes the major signaling pathways related to HCC, such as the PI3K/AKT/mTOR signaling pathway, RAS/RAF/MEK/ERK signaling pathway, and VEGF/VEGFR, PDGFR, and FGFR signaling pathway. New advances in the corresponding molecular targeted therapy for HCC are described, and the perspectives on future direction of relevant research are discussed.

Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease characterized by excessive fat accumulation in the liver. More and more evidence suggests that NAFLD is a multisystem disease that affects multiple extra-hepatic organs. Recent studies have shown that NAFLD may be associated with the risk, severity, and outcome of ischemic stroke. The article provides a summary of these aspects.

Objective To explore the antioxidant mechanism ofhistone deacetylase 2 (HDAC2) regulating Nrf 2 acetylation in lipopolysaccharide (LPS)-induced type Ⅱ alveolar epithelial cell injury.Methods The experiment was divided into two parts.The first part was the routine culture of type Ⅱ alveolar epithelial cells of mice.The cells were stimulated with different concentrations of LPS (10 ng/ mL,100 ng/mL and 1 000 ng/mL).CCK-8 was used to detect the cell activity at 0 h,6 h,12 h,24 h and 48 h,respectively.The second part:Alveolar epithelial cells of type Ⅱ were cultured and divided into the normal control group (control group),LPS group,HDAC2 lentivirus interference group (siRNA-HDAC2 group) and HDAC2 lentivirus overexpression group (LV-HDAC2 group).The expression of HDAC2 and Nrf2 were detected by Western blot,the acetylation of Nrf2 was detected by immunoprecipitation,and the stability of nrf2 was detected after actinidone action.The activity of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by chemical colorimetry.SPSS 23.0 statistical software was used.LSD-t test was used for comparison between two groups,and one-way ANOVA test was used for comparison among multiple groups.Results Compared with the control group,the expression of HDAC2 protein in the LPS group increased (t=5.974,P=0.027),the acetylation level of Nrf2 decreased (t=7.223,P=0.002),the Nrf2 protein level increased (t=2.929,P=0.043),the protein stability of Nrf2 increased,the SOD activity decreased (t=121,P＜0.01),and the MDA content increased (t=10.45,P=0.000 5).Compared with the LPS group,Nrf2 acetylation level decreased in the LV-HDAC2 group (t=1 1.29,P=0.000 4),Nrf2 protein expression increased (t=3.194,P=0.033),Nrf2 protein stability increased,SOD activity increased (t=4.678,P=0.009),and MDA content decreased in the LV-HDAC2 group (t=5.417,P=0.005 6).While the opposite trend was observed in the siRNA-HDAC2 group.Conclusion After LPS stimulation,oxidative stress of type Ⅱ alveolar epithelial cells was aggravated.HDAC2 could decrease the level of Nrf2 acetylation,increase the expression of Nrf2 protein,and alleviate LPS-induced oxidative stress.

Objective:To establish a new bile duct injury and repair model in mice by generating bile duct distal stricture and proximal dilatation.Methods:The mice were randomly divided into sham operation group, bile duct stricture (BDS) group and bile duct ligation (BDL) group. The dilated bile duct of BDS mice was injured and then repaired 14 days after the modeling operation. Biochemical markers were detected and histopathological changes were analyzed.Results:14 days after the establishment of the model, the body mass in BDL group was significantly lower than that of the sham group ( P<0.05), while the body mass in BDS group was similar to sham group. Compared with the sham group, the bile duct and gallbladder of the BDS group and BDL group were both prominently dilated, but the sum of the diameters of bile duct and gallbladder in BDS group was significantly smaller than that in the BDL group ( P<0.05). Indocyanine green fluorescence imaging confirmed that biliary tract of BDS group could still drain bile. Serum ALT, AST and TBil levels in the BDS group were slightly higher than those in the sham group (all P<0.05), but significantly lower than those in the BDL group ( P<0.05). Bile ducts of BDS mice were injured by notching and repaired with bile duct path. 30 days after the repairing, HE staining showed that the bile duct epithelium around the patch was arranged in orderliness. Immunohistochemistry confirmed the positive staining of green fluorescent protein (EGFP) and CK19 in those groups. Conclusion:This model of bile duct injury and repair in mice can provide a new model for the study of the mechanism of bile duct injury and repair and the evaluation of tissue engineering bile duct.

Objective To explore the diagnosis of clinically suspicious von Willebrand disease(vWD)in a family and its pathogene-sis.Methods The pedigree information and the biological specimen were collected from the clinically suspected VWD patient and her family members(4 persons in total)in Peking University First Hospital.The levels of platelet count(PLT),activated partial thrombo-plastin time(APTT),vWF antigen(vWF:Ag),vWF activity(vWF:Ac)and FⅧ activity(FⅧ:C)were detected,and vWF risto-cetin cofactor(vWF:RCo)assay,ristocetin-induced platelet aggregation assay(RIPA)and vWF collagen binding(vWF:CB)assay were performed for phenotype diagnosis.The peripheral blood genomic DNAs were extracted from the proband and her family members to perform whole-exome sequencing for identifying the mutation of vWF gene,The mutation site was analyzed by using bioinformation tools to explore the pathogenesis of the proband.Results The APTT of proband(m 1)was slightly prolonged and her vWF:Ag,vWF:Ac,vWF:RCo and vWF:CB were significantly decreased.There was no obvious aggregation in RIPA assay(1.0 mg/mL and 1.25 mg/mL).In her father(Ⅱ3),APTT,FⅧ:C,vWF:Ag,vWF:Ac and vWF:CB were normal,but vWF:RCo was slightly decreased.In her mother(Ⅱ4),APTT,FⅧ:C,vWF:Ag,vWF:RCo and vWF:CB were all normal,but vWF:Ac significantly decreased.In her brother(Ⅲ2),APTT and FⅧ:C were normal,but vWF:Ag,vWF:Ac,vWF:RCo and vWF:CB were reduced to varying degrees.In all the family members(father,mother and brpther),no apparent aggregation in RIPA(1.0 mg/mL)was shown.Genetic analysis showed that the proband(Ⅲ1)carried a compound heterozygous mutation of vWF gene c.7288-9T＞G and c.1117C＞T,her father(Ⅱ3)carried vWF gene c.7288-9T＞G heterozygous mutation,and vWF gene c.1117C＞T heterozygous mutation was presented in both mother(Ⅱ4)and brother(Ⅲ2).Conclusion According to the results of laboratory tests,the proband was diagnosed as type 2A vWD.The hetero-zygous mutation in vWF gene c.1117C＞T and c.7288-9T＞G may be the molecular mechanism leading to type 2A vWD in the proband.

With the proposal and development of new technologies, new methods, and new education theories, the research on medical education and teaching has been conducted extensively. The theories, methods, and management mode of the research on medical education and teaching are keeping pace with the times and thus promote the rapid improvement in teaching ability, medical talents, and the training level of medical humanities. However, current studies on medical education and teaching focus too much on the innovation of education theories, methods, and management and neglect the aspects such as educational ethics, student awareness, and assessment of potential benefits and risks. In addition, the lack of unified implementation and supervision standards in the whole process management of education research is also a problem that needs to be solved urgently, especially in empirical studies, which brings great challenges to the standardized implementation of education research. By performing an analogy with the clinical trial research system and summarizing education research and educational ethics, this study, for the first time, proposes a set of standardized and systematic medical education and research standards, including the paradigms of classification, staging and different stages. These standards provide a theoretical basis for implementing medical education research, improving the education compliance of target students, fully guaranteeing the benefits of target students, and avoiding the risks in education research, which provides a reference for standardized education research in other disciplines.

Objective:To explore effects of impaired glucose tolerance (IGT) and type 2 diabetes mellitus (T2DM) on the 5-year prognosis after percutaneous coronary intervention (PCI) for coronary heart disease.Methods:A total of 165 patients receiving PCI from December 2011 to September 2013 were selected by the convenient sampling method. According to the results of oral glucose tolerance test, they were divided into T2DM group (54 cases) , IGT group (52 cases) , and non-diabetes mellitus (NDM) group (59 cases) . The patients were followed up in the outpatient clinic after discharge and recorded the occurrence of major adverse cardiovascular events (MACE) of patients within 5 years after PCI, including cardiogenic death, non-fatal reinfarction, non-fatal stroke and re-hospitalization for heart failure.Results:The 5-year follow-up results showed that 72 patients had MACE, of which 15 died of cardiogenic death, 11 were admitted to the hospital due to heart failure, 5 had non-fatal stroke and 41 had non-fatal reinfarction. Among the 72 patients with MACE, there were 17 cases in the NDM group, 30 cases in the IGT group and 25 cases in the T2DM group. The IGT group was higher than the NDM group, and the difference was statistically significant ( P<0.05) , but there was no statistically significant difference between the IGT group and the T2DM group ( P>0.05) . Among the 41 non-fatal reinfarction patients, there were 10 cases in the NDM group, 19 cases in the IGT group and 12 cases in the T2DM group. The IGT group was higher than the NDM group and the T2DM group, and the differences were statistically significant ( P<0.05) . Conclusions:IGT increases the risk of myocardial infarction in patients after PCI, which is a risk factor for MACE events.