Objective:To clone human anti-ricin antibodies from large phage antibody library.Methods:Panning for a large phage library against ricin toxin was conducted to select specific antibodies against ricin. The binding activities and specificities were tested by ELISA method. Soluble ScFvs were prepared through infecting E coli. HB2151 with the selected phage antibodies and induction with IPTG. Results:Forty positive clones were obtained after 5 rounds of panning, and 12 clones had specific binding ability to ricin toxin. DNA fingerprinting showed 7 different band patterns indicating 7 different positive clones. DNA sequencing showed that variable regions of these ScFvs belonged to different subgroups.Conclusion:Human anti-ricin antibodies were successfully obtained from large phage antibody library.

Objective To explore nurses' bonus assessment and distribution methods based on the information technology platform and to improve the efficiency of nursing management. Methods We build a hospital informa-tion platform in order to collect nursing work efficiency data by using existing computer information network. We distributed the bonus through the information platform according to evaluate nursing quality and performance effi-ciency. Results After the information flatform used for 2 years,the satisfaction of nurses with different professional titles and different experience showed no significant difference (P>0.05). The time of department management person-nel collecting and distributing data change from [103.75 (180.00,215.00)] min per month to [42.50 (60.00,90.00)] min per month.The difference showed statistical significance(P<0.05). Conclusion The technology platform can im-prove the satisfaction and the nursing management efficiency.

Objective To screen and verify the proteins interacting with phosphorylation cluster of DNA dependent protein kinase catalytic subunit((DNA-PKcs) by yeast two-hybrid assay.Methods To know the proteins interacting with DNA-PKcs phosphorylation cluster,yeast two-hybrid assay was applied to screen the cDNA library of human hepatic tissue with a previously constructed plasmid pGBKT7-DPC.The positive clones were further identified by PCR,rotary validation and sequence analysis.Then the eukaryotic expression vectors of the bait protein and screened positive clone proteins were constructed and transfected into human embryonic kidney 293T cells to detect whether the proteins could been expressed correctly.At last,the bait protein and screened positive clone proteins were co-transfected into 293T cells and protein interaction was detected with Co-Immunoprecipitation (Co-IP) assay.Results After two rounds of screening using the yeast two-hybrid assay,12 candidate clones were obtained.Then 7 clones with different insert fragments were identified by PCR,and 3 positive proteins interacted with DNA-PKcs phosphorylation cluster were further verified by rotary validation.Sequencing analysis demonstrated that these 3 proteins were MBNL1,SIK2 and YY1AP1,respectively.Accordingly,the eukaryotic expression vectors of bait protein and 3 positive clone proteins were constructed successfully and expressed correctly in 293T ceils.Finally,the Co-IP assay confirmed that these 3 positive clone proteins could interact with DNA-PKcs phosphorylation cluster.Conclusions Proteins interacting with DNA-PKcs phosphorylation cluster are successfully screened and identified.

Objective To explore the predictive value of ventricle late potential (LP) for arrhythmic events in the patients with Brugada syndrome. Methods Totally 43 patients with Brugada syndrome were divided into symptom group (n=24) and asymptom group (n=19).Signalaveraged electrocardiography(SAECG) was performed to analyze characteristics of LP in all subjects.The occurrence of arrhythmic events was observed in all patients during the dynamic follow-up for (33.8±9.0) months.Results There were 22 cases (91.7％) and 7 cases (36.8％) with LP positive in patients with symptom and asymptom,respectively.The incidences of arrhythmic events were 72.4％ in Brugada syndrome patients with positive LP and 14.3％ in patients with negative LP,respectively.The relative risk (RR,95％ CI) for LP prediction of the arrhythmic events was [5.1,(1.4～ 18.6)] (P =0.002). ConclusionsLP may be one of effective factors predicting arrhythmic events in the patients with Brugada syndrome.

Objective: To understand the prevalence and risk factors for functional constipation (FC) among high school students in Chongming District of Shanghai. Methods: A cross sectional study was conducted in Chongming District of Shanghai from March-June 2019. A total of 4 969 adolescents under the age of 18 were recruited high schools, using a stratified random sampling technique. A validated self administered questionnaire on Rome IV criteria for diagnosing FC and predisposing factors was filled by each student in a classroom setting. Results: The prevalence of FC among middle school students in Chongming District of Shanghai was 13.95%. There were no significant differences in the prevalence between males and females, middle and high school groups, and urban and rural areas( P ＞0.05). In senior high schools, students in the graduation year were more likely to suffer from FC(17.36%，130/749) than other students (13.77%，201/1 460)( χ 2=5.01, P =0.03). The prevalence of FC in the key senior high schools(18.23%，115/631) was significantly higher than that in ordinary high schools (13.07%，49/375)( χ 2=7.43, P =0.02). Multivariate Logistic regression analysis showed that frequency of physical exercise, and consumption of spicy foods, proportion of spicy foods in the diet, consumption of vegetables, a lower proportion of vegetables in the diet, drinking water, anorexia, quality of sleep and school type were associated with FC in high school students ( OR=0.11-7.71, P <0.05). Conclusion FC is prevalent among high school students on Chongming District of Shanghai, especially among middle school graduates, and many risk factors were significantly associated with the occurrence of FC.

A 69-year old female patient was admitted because of 3 days of worsened chest pain.Coronary angiography showed60% stenosis of distal left main stem,chronic total occlusion of left anterior descending (LAD),70% stenosis at the ostium of a smallleft circumflex,70-90%stenosis at the paroxysmal and middle part of a dominant fight coronary artery (RCA),and a normal left internalmammary artery (LIMA) with normal origination and orientation.Percutaneous intervention was attempted but failed on the occludedlesion of LAD.The patient received minimally invasive direct coronary artery bypass (MIDCAB) with left LIMA isolation by Davincirobot.Eleven days later,the RCA lesion was treated by Sirolimus Rapamicin eluting stents implantation percutaneously.Then thepatient was discharged uneventfully after 3 days hospitalization.Our experience suggests that two stop shops of hybrid technique befeasible and safe in the treatment of elderly patient with multiple coronary diseases.

Background: The detection of glutamic acid decarboxylase 65 (GAD65) autoantibodies is essential for the prediction and diagnosis of latent autoimmune diabetes in adults (LADA). The aim of the current study was to compare a newly developed electrochemiluminescence (ECL)-GAD65 antibody assay with the established radiobinding assay, and to explore whether the new assay could be used to define LADA more precisely. Methods: Serum samples were harvested from 141 patients with LADA, 95 with type 1 diabetes mellitus, and 99 with type 2 diabetes mellitus, and tested for GAD65 autoantibodies using both the radiobinding assay and ECL assay. A glutamic acid decarboxylase antibodies (GADA) competition assay was also performed to assess antibody affinity. Furthermore, the clinical features of these patients were compared. Results: Eighty-eight out of 141 serum samples (62.4%) from LADA patients were GAD65 antibody-positive by ECL assay. Compared with ECL-GAD65 antibody-negative patients, ECL-GAD65 antibody-positive patients were leaner (P<0.0001), had poorer β-cell function (P<0.05), and were more likely to have other diabetes-associated autoantibodies. The β-cell function of ECLGAD65 antibody-positive patients was similar to that of type 1 diabetes mellitus patients, whereas ECL-GAD65 antibody-negative patients were more similar to type 2 diabetes mellitus patients. Conclusion Patients with ECL-GAD65 antibody-negative share a similar phenotype with type 2 diabetes mellitus patients, whereas patients with ECL-GAD65 antibody-positive resemble those with type 1 diabetes mellitus. Thus, the detection of GADA using ECL may help to identify the subtype of LADA.

Background: No currently available biomarkers or treatment regimens fully meet therapeutic needs of type 1 diabetes mellitus (T1DM). Circular RNA (circRNA) is a recently identified class of stable noncoding RNA that have been documented as potential biomarkers for various diseases. Our objective was to identify and analyze plasma circRNAs altered in T1DM. Methods: We used microarray to screen differentially expressed plasma circRNAs in patients with new onset T1DM (n=3) and age-/gender-matched healthy controls (n=3). Then, we selected six candidates with highest fold-change and validated them by quantitative real-time polymerase chain reaction in independent human cohort samples (n=12). Bioinformatic tools were adopted to predict putative microRNAs (miRNAs) sponged by these validated circRNAs and their downstream messenger RNAs (mRNAs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to gain further insights into T1DM pathogenesis. Results: We identified 68 differentially expressed circRNAs, with 61 and seven being up- and downregulated respectively. Four of the six selected candidates were successfully validated. Curations of their predicted interacting miRNAs revealed critical roles in inflammation and pathogenesis of autoimmune disorders. Functional relations were visualized by a circRNA-miRNA-mRNA network. GO and KEGG analyses identified multiple inflammation-related processes that could be potentially associated with T1DM pathogenesis, including cytokine-cytokine receptor interaction, inflammatory mediator regulation of transient receptor potential channels and leukocyte activation involved in immune response. Conclusion Our study report, for the first time, a profile of differentially expressed plasma circRNAs in new onset T1DM. Further in silico annotations and bioinformatics analyses supported future application of circRNAs as novel biomarkers of T1DM.

No currently available biomarkers or treatment regimens fully meet therapeutic needs of type 1 diabetes mellitus (T1DM). Circular RNA (circRNA) is a recently identified class of stable noncoding RNA that have been documented as potential biomarkers for various diseases. Our objective was to identify and analyze plasma circRNAs altered in T1DM.

Objective:To explore the association between serum high density lipoprotein subtype 3 cholesterol (HDL3-C) levels and the severity and in-stent restenosis of patients with coronary artery disease.Methods:124 patients with coronary artery diseases and 62 healthy controls were included in this clinical case-control retrospective study. Participants were hospitalized from November 2020 to November 2021 at Jinling Hospital, Medical School of Nanjing University were enrolled. Patients with coronary artery disease were as follows: 28 patients with acute coronary syndrome and 96 patients with stable coronary heart disease. Serum HDL3-C levels as well as total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) levels were determined. According to the coronary artery angiography results of all patients at the time of admission, Gensini scores were calculated and patients were divided into in-stent restenosis group ( n=22), no in-stent stenosis group ( n=23) and non-stent implantation group ( n=79). The correlation between HDL3-C levels and other parameters was analyzed by Pearson or Spearman correlation analyses. Multivariate Logistic regression analyses were used to determine the impact of HDL3-C on the in-stent restenosis of coronary artery diseases. Results:Compared with controls, serum levels of HDL3-C and HDL-C were significantly decreased in patients with coronary artery diseases (all P<0.05). There was a significantly negative correlation between HDL3-C levels and Gensini scores ( r=-0.201, P=0.043). Among patients with coronary artery disease, serum levels of HDL3C, TC and TG in the in-stent restenosis group were significantly lower than in no in-stent stenosis group as well as than in the non-stent implantation group (all P<0.05). Multivariate Logistic regression analyses showed that after adjusting for age, sex, lipid-lowering drugs and TC, TG, LDLC parameters, HDL3-C ( OR=0.885, 95% CI 0.791-0.990, P=0.033) and HDL-C ( OR=0.018, 95% CI 0.001-0.426, P=0.013) levels were both independently associated with the occurrence of coronary artery disease; only HDL3-C levels (no in-stent stenosis group as the reference: OR=0.833, 95% CI 0.698-0.994, P=0.042; non-stent implantation group as the reference: OR=0.812, 95% CI 0.685-0.963, P=0.017) were independently associated with the presence of in-stent restenosis ( P<0.05). Conclusions:Serum HDL3-C levels are decreased in patients with coronary artery disease, especially in patients with in-stent restenosis. HDL3-C levels are associated with the severity of coronary artery lesions and the presence of in-stent restenosis of coronary arteries.