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Objective: Detection of gene rearrangements in MYC (a family of regulator genes and proto-oncogenes) and human B-cell lymphoma 6 (BCL6) using fluorescence in situ hybridization (FISH) are important in the evaluation of lymphomas, in particular diffuse large B-cell lymphoma (DLBCL) and Burkitt lymphoma. Our current clinical MYC and BCL6 FISH workflow involves an overnight hybridization of probes with digital analysis using the GenASIs Scan and Analysis instrument (Applied Spectral Imaging). In order to improve assay turnaround time SureFISH probes were validated to reduce the hybridization time from 16 hours down to 1.5 hours. Methods: Validation was a four-phase process involving initial development of the assays by testing new probes in a manual protocol, and cytogenetic studies to confirm the probe specificity, sensitivity, and localization. In the next phase, the assays were validated as a manual assay. The third phase involved development of the digital FISH assays by testing and optimizing the GenASIs Scan and Analysis instrument. In the final phase, the digital FISH assays were validated. Results: Cytogenetic studies confirmed 100% probe sensitivity/specificity, and localization patterns. Negative reference range cutoffs calculated from 20 normal lymph nodes using the inverse of the beta cumulative probability density function (Excel BETAINV calculation) were 11% inclusive for both manual and digital MYC and BCL6 assays. There was 100% concordance between the manual and digital methods. The shortened hybridization time decreased the overall workflow time by 14.5 hours. Conclusions: This study validates the use of the SureFISH MYC and BCL6 probes on formalin fixed paraffin embedded (FFPE) tissue sections using a hybridization time of 1.5 hours that shortened the overall workflow by 14.5 hours. The process described also provides a standardized framework for validating digital FISH assays in the future.

AimsThe study was designed to develop a measurement for the motivation for and against change in methamphetamine users in the compulsory detoxification setting.DesignThis is a cross-sectional study.SettingThe study was carried out in a compulsory detoxification center for male drug users in China.ParticipantsA total of 228 male methamphetamine users who had undergone the program for at least 30 days.MeasurementsThe motivation for/against change relating to compulsory detoxification was carried out using the Likert scale. A series of questionnaires were filled out by the participants, including the Egna Minnen Beträffande Uppfostran for rearing style, the Barratt Impulsiveness Scale-11, the adult ADHD self-report scale, and the Pittsburgh sleep quality index. Participants were also asked to recall the withdrawal symptoms before the program and to rate their current craving levels.FindingsMotivations were grouped into three factors, namely, the expectation to use drugs upon the completion of the program (factor 1), the disagreement with the compulsory setting (factor 2), and the motivation to quit drug use (factor 3). Cronbach's alpha values were 0.8037, 0.8049, and 0.6292, respectively. The structural equation model showed that the overall motivation was characterized by motivation against change rather than that for change. The overall motivation was also directly affected by the current craving level and indirectly affected by the severity of addiction, paternal authoritarian upbringing style, and ADHD traits.ConclusionThis study provided a measurement of motivation for and against change in subjects with drug misconduct and suggested that the motivation against change may disclose more psychological barriers than the motivation for change.

OBJECTIVE: To investigate the efficacy and mechanism of Qifu Lizhong enema prescription(芪附理中灌肠方, QFLZ) on intervening ulcerative colitis (UC) rat model with TCM spleen and kidney Yang insufficiency syndrome. METHODS: Seventy-two male Sprague-Dawley rats were randomly assigned to six groups: normal model, mesalazine, and QFLZ high, medium, and low dose groups, each with 12 rats. After 3 d of adaptation feeding, all groups except the normal group were induced using rhubarb decoction in combination with trinitrobenzene sulfonic acid (TNBS)/55 % ethanol to establish a UC rat model. Following successful modeling, the normal and model groups received daily saline enema, while the Chinese medicine and Western medicine groups received daily QFLZ and Mesalazine enema for 2 weeks respectively. The disease activity index score, hematoxylin and eosin staining, immunohistochemistry, and Western blotting were used to determine the expression of claudin 1, claudin 2, zonula occludens-1 protein (ZO-1), and F-actin proteins in each rat colon tissue following treatment. RESULTS: QFLZ significantly alleviated the structural disorganization in the form of epithelial glands in the intestinal mucosa of rats with UC and retarded the progression of the disease. The intestinal mucosal epithelial cells of UC rats showed decreased expression of claudin 1, ZO-1, F-actin (P < 0.05), claudin 2 appeared elevated (P < 0.05), which resulted in impaired TJ. Treatment with QFLZ resulted in elevated expression of claudin 1 (P < 0.05), ZO-1 (P < 0.05) and F-actin (P < 0.05) and decreased expression of claudin 2 (P < 0.05), which allowed for repair of the intestinal mucosal TJ, which in turn served as a treatment for UC. CONCLUSIONS: The mechanism of repairing TJ function and repairing the intestinal mucosal barrier by QFLZ may be associated with up-regulation of claudin 1, ZO-1, and F-actin levels, and down-regulation of claudin 2 expression level.

Background For emergency department (ED) patients, risk assessment, prophylaxis, early diagnosis and appropriate treatment of venous thromboembolism (VTE) are essential for preventing morbidity and mortality. This study aimes to investigate knowledge amongst emergency medical staff in the management of VTE. Methods We designed a questionnaire based on multiple scales. The questionnaire was distributed to the medical and nursing clinical staff in the large urban ED of a medical center in Northern China. Data was described with percentages and the Kruskal-Wallis test was used to compare ranked data between different groups. The statistical analysis was done using the SPSS 22.0 software. Results In this survey, 180 questionnaires were distributed and 174 valid responses (response rate of 96.67%) were collected and analyzed. In scores of VTE knowledge, no significant differences were found with respect to job (doctor vs. nurse), the number of years working in clinical medicine, education level, and current position, previous hospital experience and nurses' current work location within the ED. However, in pair wise comparison, we found participants who worked in ED for more than 5 years (n=83) scored significantly higher on the questionnaire than those under 5 years (n=91) (95.75 vs. 79.97, P=0.039). There was a significant difference in some questions based on gender, age, job, and nurse work location, number of working years, education level, and different ED working lifetime. Conclusion Our survey has shown deficiencies among ED medical staff in knowledge and awareness of the management of VTE. We recommend several changes be considered, such as the introduction of an interdisciplinary workshop for medical staff; the introduction of a standardized VTE protocol; a mandatory study module on VTE for new physicians and nurses; the introduction of a mandatory reporting system for adverse events (including VTE).

Objective(s): MiRNAs have been proposed to be key regulators of tumorigenesis, progression and metastasis. However, their effect and prognostic value in gastric cancer is still poorly known. Materials and Methods: Gastric cancer cell lines were cultured. Tissue samples obtained from 36 gastric cancer patients were used for quantitative real-time PCR (qRT-PCR) analysis. The tissue microarrays (TMAs) consisted of 126 cases of gastric carcinoma that were used for In situ hybridisation (ISH). Lentivirus plasmids were co-transfected into 293FT cells. Cell migration was examined using wound-healing assays. Statistical analyses were performed using SPSS16.0 software. Results: In this study, we found that the expression levels of miR-493 were strongly down-regulated in gastric cancer and were associated with clinical stage and the presence of lymph node metastases. Moreover, miR-493 might independently predict OS and RFS in gastric cancer. We further found that up-regulation of miR-493 inhibited the proliferation and metastasis of gastric cancer cells, in vitro and in vivo. In addition, miR-493 directly targeted RhoC, which resulted in a marked reduction of the expression of mRNA and protein. This effect, in turn, led to a decreased ability of growth, invasion and metastasis in gastric cancer cells. Conclusion: Taken together, our findings demonstrate that miR-493 is important for gastric cancer initiation and progression and holds promise as a prognostic biomarker to predict survival and relapse in gastric cancer. It is also a potential therapeutic tool to improve clinical outcomes in this disease.

Evidence suggests short or long sleep duration is associated with a higher risk of diabetes. Using a large longitudinal data set spanning 2 years, we examined whether a change in sleep duration is associated with diabetes. Current analysis included 56,588 participants who were free of diabetes during both 2006–2007 (exam1) and 2008–2009 (exam2). Sleep duration was categorized into 7 groups: ≤5.5 hours, 6.0 to 6.5 hours, 7.0 hours, 7.5 to 8.0 hours, ≥8.5 hours, decrease ≥2 hours, and increase ≥2 hours. Cox proportional hazards models were used to calculate hazard ratios (HRs) and their confidence intervals (CI) for diabetes, according to sleep duration. Compared to the reference group of persistent 7-h sleepers, participants who slept 7.5 to 8 hours per night (HR, 1.20; 95% CI, 1.02–1.40), ≥8.5 hours per night (HR, 1.37; 95% CI, 1.03–1.81) and an increase of ≥2 hours sleep per night (HR, 1.24; 95% CI, 1.05–1.48) were all associated with an increased risk of developing diabetes in analyses adjusted for age, sex, education level, income level, smoking status, drinking status, physical activity, BMI, snoring status, hypertension, hyperlipidemia, and family history of diabetes. The abovementioned associations of sleep duration and incident diabetes were only prominent among individuals aged

Ecological city (eco-city) and its planning approach are emerging concepts in urban study, urban planning, ecological economics, environmental policy and corporate environmental management. However, opinions remain divided over the connotation and denotation of the term “ecological city”, what key issues ecocity planning can solve, and its specific contents. In this study, we present 10 basic propositions that define the eco-city and clarify its key parameters, thereby providing the basis for discussing the assumptions and principles underlying different approaches to sustainable urban development. We then summarize the concept and principles of an eco-city, and define the main requirements for ecocity planning. We conclude that an eco-city is a city in which the urban population, scale of land use and intensity of human activities are limited to the regional resource and environmental carrying capacity, which does not cause increasing or irreversible damage to the regional ecosystem’s structure,...

G-quadruplexes (G4) are four-stranded DNA secondary structures, which are involved in a diverse range of biological processes. Although the anti-cancer potential of G4s in promoters of oncogenes has been thoroughly investigated, the functions of promoter G4s in non-cancer-related genes are not well understood. The aims of this thesis are (i) to investigate the biological importance of promoter G4s in cardiac function-related genes, and (ii) to explore the feasibility and potential of multitargeting strategy at the single gene level. These aims have been achieved by revealing the prevalence of G4s in TRRs (transcription regulatory regions) of cardiac function-related genes via bioinformatics approaches, and then investigating potential biological significance of G4s in promoters of human MEF2D and TnIc genes via a combination of biophysical, molecular, and cell biology approaches. By using bioinformatics approaches, TRRs of cardiac function-related genes were found to be enriched by potential G4-forming sequences. According to these results, G4s from the promoters of human MEF2D and TnIc were chosen and subjected to biophysical characterisations in solution. By using EMSA, DMS footprinting, CD and smFRET spectroscopy, the formation, thermodynamic stability, and unfolding kinetics of MEF2D and TnIc G4s were investigated. Briefly, in 100 mM K+, the MEF2DG4 can adopt a hybrid of very stable parallel/anti-parallel G4(s), while the TnIc MNSG4 and -80G4 are able to adopt anti-parallel and parallel structures, respectively, with comparable stability as compared to other oncogene G4s. Subsequently, cooperative regulatory roles of TnIc G4s as enhancers once stabilised by a G4-binding ligand (di-copper complex) were observed in vitro mainly by dual luciferase reporter assays in HEK293 cells. Three possible G4-mediated mechanisms have been proposed. This work provides the first indication about the biological significance of promoter G4s in cardiac function-related genes, and the feasibility to target multiple G4s at the single gene level.

ADAMTS13 (a disintegrin and metalloprotease with thrombospondin motifs), a circulating multidomain zinc metalloprotease of the reprolysin subfamily, is critical for preventing von Wille-brand factor-platelet interaction under high shear stress conditions. A deficiency of the protease, due to mutations in the ADAMTS13 gene or the presence of antibodies that inhibit the activity of the protease, causes thrombotic thrombocytopenic purpura (TTP). Plasma therapy, the conventional therapy for TTP, may cause serious adverse reactions and is ineffective in some patients. In order to develop new strategies for improving the diagnosis and treatment of TTP, we produced a series of truncated ADAMTS13 proteins in mammalian cells and analyzed their binding with and suppression by the IgG derived from the TTP patients. The results revealed that truncation of the ADAMTS13 protein at its cysteine-rich region eliminated its recognition by the antibodies without abolishing its von Willebrand factor-cleaving activity. This raises the possibility that resistant ADAMTS13 variants may be exploited to circumvent inhibitory antibodies that cause TTP.

The effectiveness of breeding Indica photoperiod-sensitive genic male-sterile (PGMS) rice has been studied since 1982. Eight Indica male sterile lines have been bred from the progenies of Indica/Japonica or its reciprocals; their fertility response to photoperiod and temperature and fertility alteration were investigated. Five promising two-line inter-varietal hybrid rices of W6154S and W6184S have been developed through extensive test-crossing. Discussions made on the evaluation and utilization of Indica PGMS or TGMS lines in current two-line hybrid rice breeding.

Objective To probe the effect of electroacupuncture (EA) at Shenshu (BL 23) on morphine withdrawal syndrome and the mechanism. Methods Morphine dependence was induced in the rat by subcutaneous injection of morphine, and the acute withdrawal syndrome was precipitated b y intraperitoneal administration of naloxone. The rats were pretreated with 100 H z EA at Shenshu (BL 23) for one hour before the naloxone challenge. Withdrawal scores were calculated, and c-fos levels were detected by immunocytochemical meth od. Results EA at Shenshu (BL 23) could obviously inhibit withdrawal syndrome, decreased expression of c-fos in the PAG, paraventricular hypothalamus nucleus, lateral hypothalamic area and hypocampus while EA increase c-fos positive neurons in the amygdula and core of nucleus accumbens. Conclu sion EA at Shenshu (BL 23) can effectively inhibit morphine withdrawal s yndrome and many central nucleus groups are involved in the inhibition of EA on opioid m edicine dependence.